ПОЛИПЕПТИД (ВАРИАНТЫ), ДЕЙСТВУЮЩИЙ В КОМПЛЕКСЕ С РЕЦЕПТОРОМ АНДРОГЕНА, НУКЛЕИНОВАЯ КИСЛОТА (ВАРИАНТЫ), ВЕКТОР (ВАРИАНТЫ) И КЛЕТКА-ХОЗЯИН (ВАРИАНТЫ)

Изобретение относится к биотехнологии и генной инженерии. Полипептид с последовательностью SEQ ID NO:3 связывается с рецептором андрогенов и увеличивает способность рецептора андрогенов трансактивировать андроген-чувствительный ген. Предложены также нуклеиновая кислота, кодирующая такой полипептид, и вектор, содержащий такую нуклеиновую кислоту. Группа изобретений может быть использована для создания трансгенных животных, экспрессирующих ген белка, действующего в комплексе с рецептором андрогена, служащих моделями для разработки лекарственных средств для лечения рака. 12 н. и 12 з.п. ф-лы.

МНОЖЕСТВЕННЫЕ ВАРИАНТЫ МЕНИНГОКОККОВОГО БЕЛКА NMB1870

Изобретение касается композиции для лечения заболевания, вызываемого Neisseria meningitides. Согласно изобретению белок NMB 1870 представляет собой эффективный антиген для получения иммунного ответа против менингококка, и он экспрессируется среди всех менингококковых серогрупп. Идентифицированы сорок две различные последовательности NMB 1870, и они образуют группы трех вариантов. Сыворотка, полученная против данного варианта, бактерицидна внутри той же самой группы варианта, но неактивна против штаммов, которые экспрессируют один из двух других вариантов, т.е. существует внутривариантная, но не межвариантная перекрестная защита. Поэтому в данном изобретении для максимальной перекрестной штаммовой эффективности используют смеси, содержащие различные варианты NMB 1870. В изобретении раскрывается способ индуцирования иммунного ответа млекопитающего, предусматривающий использование композиций на основе различных вариантов NMB 1870, а также представлены нуклеиновые кислоты, кодирующие указанные ...

НОВЫЕ БИЦИКЛИЧЕСКИЕ НУКЛЕОЗИДЫ И ОЛИГОМЕРЫ, ПОЛУЧЕННЫЕ ИЗ НИХ

Изобретение относится к новым соединениям формулы (I) и олигонуклеотидам, полученным из них. В частности, настоящее изобретение относится к соединению формулы (I) где один из T1 и Т2 представляет собой OR1 или OR2; и другой из T1 и Т2 представляет собой OR1 или OR2; где R1 представляет собой Н или гидроксил-защитную группу, где указанная гидроксил-защитная группа независимо в каждом случае выбрана из ацетила, бензила, трет-бутилдиметилсилила, трет-бутилдифенилсилила, тритила, 4-монометокситритила, 4,4'-диметокситритила (DMTr), 4,4',4''-триметокситритила (TMTr), 9-фенилксантин-9-ила (пиксила) и 9-(п-метоксифенил)ксантин-9-ила (МОХ), и R2 представляет собой фосфорсодержащий фрагмент, где указанный фосфорсодержащий фрагмент представляет собой фосфорамидитный фрагмент, представленный формулой (X) где R5 представляет собой С1-С9 алкил, замещенный циано; R6 и R7 независимо представляют собой С1-С9 алкил; или совместно с атомом азота, к которому они присоединены, образуют гетероциклическое кольцо ...

НАБОРЫ И СПОСОБ ДЕТЕКТИРОВАНИЯ ПАПИЛЛОМАВИРУСА ЧЕЛОВЕКА С ПОМОЩЬЮ НАБОРА ОЛИГОНУКЛЕОТИДНЫХ ГРАНУЛ

Изобретение относится к области биотехнологии, в частности к набору праймеров для амплификации гена L1 папилломавируса человека (HPV), набору для детектирования генотипа папилломавируса человека (HPV) и к способам детектирования генотипов папилломавируса человека (HPV). Способ включает осуществление первичной ПЦР-амплификации гена HPV L1 в анализируемом образце с использованием набора из прямого и обратного праймеров, специфичных для участка гена L1. После чего осуществляют вторичную ПЦР-амплификацию продукта первичной ПЦР-амплификации гена L1 с использованием обратного праймера, с получением меченого биотином одноцепочечного гена L1. Затем проводят реакцию гибридизации продукта вторичной ПЦР-амплификации, меченого биотином одноцепочечного гена L1 с одним или более зондов для детектирования HPV-генотипа или зонда для детектирования HPV-генотипа. Далее осуществляют проведение реакции продукта реакции гибридизации с фикоэритрином, связанным со стрептавидином. Затем проводят измерение уровня ...

ДНК, КОДИРУЮЩАЯ МОДИФИЦИРОВАННОЕ АНТИТЕЛО ИЛИ СОЕДИНЕНИЕ С АКТИВНОСТЬЮ АГОНИСТА ТРО, СПОСОБ ИХ ПОЛУЧЕНИЯ И ЖИВОТНАЯ КЛЕТКА ИЛИ МИКРООРГАНИЗМ, ИХ ПРОДУЦИРУЮЩИЕ

Изобретение относится к ДНК, кодирующей модифицированное антитело, способное распознавать и поперечно сшивать рецептор ТРО и содержащее две или более V-области Н-цепи и две или более V-области L-цепи исходного антитела, соединенные напрямую или через линкер ковалентной или нековалентной связью, с меньшим размером по сравнению с исходным антителом. ДНК включает по две или более ДНК-последовательности, кодирующие V-области L-цепи и Н-цепи, при этом ДНК, кодирующая по меньшей мере одну из V-областей L-цепи и/или Н-цепи включает определенные нуклеотидные последовательности, приведенные в описании. В изобретении раскрыта ДНК, кодирующая соединение, проявляющее равное или большее агонистическое действие (ED50) сравнительно с тромбопоэтином (ТРО), при этом соединение может представлять собой модифицированное антитело, целое антитело или F(ab')2, способного специфически распознавать и поперечно сшивать рецептор ТРО. Изобретение также относится к вектору, содержащему указанную ДНК, животной клетке ...

СПОСОБ ВЫПОЛНЕНИЯ МАНИПУЛЯЦИИ С ПОСЛЕДОВАТЕЛЬНОСТЬЮ НУКЛЕИНОВОЙ КИСЛОТЫ, УСТРОЙСТВО ДЛЯ ПРОВЕДЕНИЯ МАНИПУЛЯЦИЙ НА ПОСЛЕДОВАТЕЛЬНОСТИ НУКЛЕИНОВОЙ КИСЛОТЫ, ПРОТОЧНЫЙ СОСУД, ТВЕРДЫЙ НОСИТЕЛЬ ДЛЯ ИММОБИЛИЗАЦИИ ПОСЛЕДОВАТЕЛЬНОСТИ НУКЛЕИНОВОЙ КИСЛОТЫ, ТВЕРДЫЙ НОСИТЕЛЬ, СПОСОБ ПОЛУЧЕНИЯ НОСИТЕЛЯ ДЛЯ ИММОБИЛИЗАЦИИ ПОСЛЕДОВАТЕЛЬНОСТИ НУКЛЕИНОВОЙ КИСЛОТЫ

Способ применим, в частности для анализа медицинских проб. Способ проведения манипуляции последовательностью нуклеиновой кислоты предусматривает обеспечение системы твердого носителя, с которым связан одноцепочечный олигонуклеотид, комплементарный специфической последовательности на нуклеиновой кислоте-мишени, более длинной, чем данный олигонуклеотид, добавление источника одноцепочечной нуклеиновой кислоты-мишени к системе твердого носителя, гибридизацию нуклеиновой кислоты-мишени с олигонуклеотидом и проведение манипуляции на гибридизованной нуклеиновой кислоте-мишени. Манипуляция может представлять собой, например, копирование или амплификацию последовательности нуклеиновой кислоты-мишени. В предпочтительном варианте обеспечен носитель в проточном сосуде, который облегчает промывание носителя для удаления примесей и оставления "чистой" пробы нуклеиновой кислоты-мишени, на которой может быть проведена манипуляция. Предпочтительно носитель имеет силоксановый матрикс, с которым связан олигонуклеотид ...

УЛУЧШЕННАЯ ОЛИГОНУКЛЕОТИДНАЯ КОНСТРУКЦИЯ ТИПА НАНОЧАСТИЦЫ, ОБЛАДАЮЩАЯ ВЫСОКОЙ ЭФФЕКТИВНОСТЬЮ, И СПОСОБ ЕЕ ПОЛУЧЕНИЯ

Изобретение относится к области биотехнологии, конкретно к самособирающейся в наночастицу олигонуклеотидной конструкции, и может быть использовано в медицине. Олигонуклеотидная конструкция согласно настоящему изобретению может быть полезной в качестве системы доставки на основе олигонуклеотида нового типа, а также инструмента для лечения злокачественных заболеваний, инфекционных заболеваний и т.п. Полимерное соединение, входящее в состав конструкции, связано с антисмысловым олигонуклеотидом посредством ковалентной связи для улучшения стабильности in vivo олигонуклеотида и эффективности его доставки в целевую клетку. В сравнении с известными аналогами заявленная олигонуклеотидная конструкция оптимизирована в отношении гомогенного материала, упрощена схема ее твердофазного синтеза, а размер двухцепочечной конструкции олиго-РНК может быть точно отрегулирован посредством контроля числа повторов блока и гидрофильного материала. 8 н. и 21 з.п. ф-лы, 18 ил., 4 табл., 6 пр.

СПОСОБ ПЕЧАТИ БИОЛОГИЧЕСКИХ ЛИГАНДОВ В ПРОЦЕССЕ ИЗГОТОВЛЕНИЯ БИОЧИПОВ

Изобретение относится к биотехнологии. Предложен способ печати биологических лигандов, представляющих собой олигосахариды, и/или полисахариды, и/или пептиды, и/или гликопептиды, и/или биотин. Предварительно на подложку наносят слой гидрофобной нелетучей жидкости, не смешивающейся с растворителем биологических лигандов и имеющей удельную массу меньше, чем удельная масса раствора биологических лигандов. Затем бесконтактным методом осуществляют печать биологических лигандов. В качестве гидрофобной нелетучей жидкости используют вазелин, минеральное масло или высшие предельные углеводороды или их смесь. Толщина слоя гидрофобной нелетучей жидкости предпочтительно составляет 50-200 мкм. Техническим результатом изобретения является повышение равномерности высыхания капель раствора биологических лигандов на подложке при одновременном обеспечении защиты биологических лигандов от преждевременного гидролиза и улучшении морфологии спотов. 2 з.п. ф-лы, 2 ил., 3 пр.

СТАБИЛЬНЫЕ И РАСТВОРИМЫЕ АНТИТЕЛА, ИНГИБИРУЮЩИЕ VEGF

Изобретение относится к области биохимии, в частности к выделенной молекуле нуклеиновой кислоты, кодирующей рекомбинантное антитело или его фрагмент, где рекомбинантное антитело или его фрагмент нейтрализует VEGF человека, а также к экспрессирующему вектору и клетке-хозяину, ее содержащей. Изобретение позволяет эффективно экспрессировать антитело или его фрагмент, которое нейтрализует VEGF человека. 3 н. и 2 з.п. ф-лы, 15 ил., 12 табл., 7 пр.

МОДУЛЯЦИЯ ЭКСПРЕССИИ HSP47

Изобретение относится к области биохимии, в частности к молекулам нуклеиновой кислоты для модуляции экспрессии генов-мишеней. Заявлена двухцепочечная молекула нуклеиновой кислоты для ингибирования экспрессии гена белка теплового шока 47 (hsp47). Также представлены композиция, включающая указанную молекулу нуклеиновой кислоты, и способ лечения пациента, страдающего от заболевания, связанного с hsp47. Изобретение может быть использовано для лечения ассоциированных с hsp47 состояний и нарушений, таких как фиброз печени, фиброз легких, фиброз брюшины и фиброз почек. 3 н. и 22 з.п. ф-лы, 27 ил., 31 табл., 10 пр.

НОВЫЕ СИНТЕТИЧЕСКИЕ АГОНИСТЫ TLR9

В изобретении описаны иммуномодулирующие соединения на основе олигонуклеотидов, которые индивидуально обеспечивают уникальные профили цитокинов и хемокинов посредством взаимодействия как агонистов TLR. Агонисты TLR9 предназначены для модуляции иммунных ответов, опосредованных Toll-подобным рецептором (TLR). Описаны также композиция, содержащая иммуномодулирующее соединение и физиологически приемлемый носитель и обеспечивающая получение иммунного ответа у пациента, способ получения иммунного ответа у пациента с использованием иммуномодулирующего соединения, способы терапевтического и профилактического лечения пациента, нуждающегося в модуляции иммунного ответа, в частности, при раке, аутоиммунном и воспалительном расстройствах, инфекционном заболевании, аллергии, астме и ряде других заболеваний. Иммуномодулирующие соединения могут использоваться в сочетании с другими терапевтическими средствами и химиотерапевтическими соединениями, используемыми при конкретных заболеваниях или расстройствах ...

КОМПОЗИЦИИ И СПОСОБЫ МОДУЛИРОВАНИЯ ЭКСПРЕССИИ HBV И TTR

Изобретение относится к применимому в медицине соединению, содержащему модифицированный олигонуклеотид и конъюгирующую группу, при этом модифицированный олигонуклеотид состоит из 12-30 связанных нуклеозидов и содержит последовательность азотистых оснований, включающую часть из по меньшей мере 8 смежных азотистых оснований, комплементарную равной по длине части SEQ ID NO: 1, при этом последовательность азотистых оснований модифицированного олигонуклеотида по меньшей мере на 90% комплементарна SEQ ID NO: 1, где конъюгирующая группа содержитгде конъюгирующая группа связана с модифицированным олигонуклеотидом на 5'-конце или 3'-конце модифицированного олигонуклеотида. Предложено новое соединение и композиция на его основе, эффективные для производства лекарственного средства для лечения заболевания, опосредованного НВV. 4 н. и 36 з.п. ф-лы, 113 пр., 108 табл.

АГОНИСТЫ РЕЦЕПТОРА КОРТИКОТРОПИН-РИЛИЗИНГ ФАКТОРА2 И ИХ ПРИМЕНЕНИЕ

Изобретение относится к биотехнологии и может быть использовано для получения агониста рецептора кортикотропин-рилизинг фактора 2 (CRF2R). Белок агониста CRF2R получают как производное CRF, урокортина I, урокортина II, урокортина III, саувагина, уротензина I и других родственных пептидов, производя аминокислотные замены в последовательностях белков. Полученный агонист CRF2R применяют в составе фармацевтической композиции для профилактики и лечения нарушений, вызываемых CRF2R, у хозяина. Изобретение позволяет разрабатывать эффективные средства для профилактики и лечения расстройств, модулируемых рецептором кортикотропин-рилизинг фактора 2, таких как мышечная дистрофия. 10 н. и 15 з.п. ф-лы, 3 табл.

СПОСОБ СИНТЕЗА НУКЛЕОЗИДНЫХ АНАЛОГОВ

Изобретение относится к способу получения 2',3'-О-алкилиден-β -нуклеозидного аналога, включающему взаимодействие 2', 3'-О-алкилиден β-фуранозилгалогенида формулы 1, где R3 и R4 представляют собой водород или (CH2)nQ, где n является целым числом от 1 до 6; Q представляют собой водород или -OR7; R5 и R6 представляют собой (С1-С6) алкил; R7 представляет собой водород или защитную группу; Y представляет хлор или бром, с гетероциклом формулы 2, где каждый из R8 представляет собой водород, фенил, фенил, замещенный галогеном, OR7 или NR7R7; R7 представляет собой водород, фенил, фенил, замещенный галогеном или защитную группу. Реакцию проводят в диметилсульфоксиде или в смеси полярных растворителей, в которой объем диметилсульфоксида составляет, по крайней мере, примерно 15%, в присутствии сильного основания, представляющего собой т-бутоксид натрия или т-бутоксид калия. Молярное отношение 2',3'-O-алкилиден-β-фуранозилгалогенида к гетероциклу составляет около (1:1) - (3:1). Молярное отношение сильного ...

ТЕРАПЕВТИЧЕСКИЕ СВЯЗЫВАЮЩИЕ МОЛЕКУЛЫ В ВИДЕ ХИМЕРНОГО АНТИТЕЛА

Изобретение относится к биотехнологии и иммунологии. Предложена связывающая молекула, представляющая собой химерное антитело к CD45RO и CD45RB. Молекула включает два домена с последовательно расположенными гипервариабельными участками CDR1, CDR2 и CDR3 и CDR1', CDR2' и CDR3'. CDR1 имеет аминокислотную последовательность NYIIH, CDR2 имеет аминокислотную последовательность YFNPYNHGTKYNEKFKG и CDR3 имеет аминокислотную последовательность SGPYAWFDT. CDR1' имеет аминокислотную последовательность RASQNIGTSIQ, CDR2' имеет аминокислотную последовательность SSSESIS и CDR3' имеет аминокислотную последовательность QQSNTWPFT. Описан соответствующий кодирующий полинуклеотид. Использование изобретения позволяет индуцировать иммуносупрессию, ингибировать Т-клеточный ответ и первичную реакцию лимфоцитов в смешанной культуре (MLR), а также удлинять время выживания мышей с тяжелым комбинированнным иммунодефицитом SCID. 2 н. и 4 з.п. ф-лы, 5 ил., 2 табл.

КОМПОЗИЦИИ И СПОСОБЫ МОДУЛИРОВАНИЯ ЭКСПРЕССИИ АПОЛИПОПРОТЕИНА C-III

Изобретение относится к пригодному для применения в медицине соединению, содержащему модифицированный олигонуклеотид и конъюгирующую группупри этом модифицированный олигонуклеотид состоит из 20 связанных азотистых оснований, комплементарных равной по длине части азотистых оснований 3533-3552 в SEQ ID NO: 3, при этом модифицированный олигонуклеотид содержит сегмент гэп, состоящий из связанных дезоксинуклеозидов; сегмент крыла 5', состоящий из связанных нуклеозидов; сегмент крыла 3', состоящий из связанных нуклеозидов; при этом сегмент гэп расположен между сегментом крыла 5' и сегментом крыла 3', и при этом каждый нуклеозид каждого сегмента крыла содержит модифицированный сахар, и где по меньшей мере один нуклеозид содержит модифицированное азотистое основание; и модифицированный олигонуклеотид содержит по меньшей мере одну модифицированную межнуклеозидную связь. Предложены новые конъюгаты модифицированных олигонуклеотидов и композиции на их основе для лечения, предупреждения или замедления ...

КОМПОЗИЦИИ И СПОСОБЫ МОДУЛИРОВАНИЯ ЭКСПРЕССИИ HBV И TTR

Группа изобретений относится к области фармакологии и медицины и направлена на лечение транстиретинового амилоидоза. Раскрывается олигомерное соединение, при этом анионная форма олигомерного соединения представлена следующей химической структурой (SEQ ID NO: 12). Кроме того, раскрыта форма свободной кислоты, натриевая соль вышеуказанного олигомерного соединения и олигомерное соединение, содержащее модифицированный олигонуклеотид и конъюгирующую группу, характеризующееся следующей формулой (SEQ ID NO: 12). Также раскрыта фармацевтическая композиция для лечения транстиретинового амилоидоза на основе вышеуказанных соединений, применение представленных соединений при производстве лекарственного средства для лечения транстиретинового амилоидоза, а кроме того, способ лечения транстиретинового амилоидоза у субъекта, включающий введение субъекту терапевтически эффективного количества олигомерного соединения или фармацевтической композиции, содержащей указанное олигомерное соединение. Группа изобретений ...

ПОЛИПЕПТИДЫ, ПРОИСХОДЯЩИЕ ИЗ ТРИПТОФАНИЛ-ТРНК-СИНТЕТАЗЫ, И ИХ ПРИМЕНЕНИЕ ДЛЯ РЕГУЛЯЦИИ РАЗВИТИЯ КРОВЕНОСНЫХ СОСУДОВ

Изобретение относится к биотехнологии. Предложены водорастворимые полипептиды (SEQ ID No.12) и (SEQ ID No.7), происходящие из полноразмерной триптофанил-тРНК-синтетазы и обладающие ангиостатической активностью в отношении неоваскуляризации глаза. Также предложены полинуклеотиды, кодирующие усеченные формы полипептидов (SEQ ID No.12) и (SEQ ID No.7), и клетка E.coli, экспрессирующая данные полипептиды. Полипептиды могут быть использованы в составе инъецируемой ангиостатической композиции и набора для ингибирования развития кровеносных сосудов глаза. Преимущество полипептидов заключается в том, что они представляют собой неиммуногенные антиангиогенные средства. 9 н. и 13 з.п. ф-лы, 5 ил., 2 табл.

Алкинсодержащий амидофосфит для функционализации синтетических олигонуклеотидов и способ его получения

Группа изобретений относится к области органической химии и химии нуклеиновых кислот и предназначена для функционализации синтетических олигонуклеотидов методом азид-алкинового циклоприсоединения («клик-химии»). Раскрывается соединение, имеющее структурную формулу, где R1 представляет собой защитную группу, стабильную в условиях синтеза олигонуклеотидов, выбранную из 4,4'-диметокситритильной (DMTr), 4-монометокситритильной (MMTr), тритильной (Tr), триметокситритильной (TMTr), 9-фенилксантен-9-ильной (Px), карбонатной или силильной групп, R2 представляет собой водород, метил или этил, X представляет собой -О- или -NH-, и L представляет собой спейсерную группу -(CH2)-n, где n = 1, 2 или 3; или -(CH2CH2O)nCH2-, где n = 1 или 2. Также раскрыт способ получения вышеуказанных амидофосфитных реагентов, включающий функционализацию карбоксильной группы линкерами, содержащими алкиновую группу, с последующей селективной защитой одной гидроксильной группы и фосфитилированием второй гидроксильной группы ...

СПОСОБ ПОЛУЧЕНИЯ 3'-ФОСФАТ,N,P-НЕЗАЩИЩЕННЫХ ФОСФОТИОАТНЫХ АНАЛОГОВ ОЛИГОДЕЗОКСИРИБОНУКЛЕОТИДОВ

Изобретение относится к области биоорганической химии, а именно к способу получения 3'-фосфат, N,P- незащищенных фосфотиоатных аналогов олигодезоксирибонуклеотидов общей формулы I, где В - остаток тимина, цитозина, аденина или гуанина; n = 1 - 20, которые могут быть использованы в качестве исходных соединений для получения фосфотиоатных олигонуклеотидных реагентов для биотехнологических целей. Способ заключается в последовательной обработке исходных защищенных олигонуклеотидов раствором йода в водном пиридине, а затем водным аммиаком и уксусной кислотой, отличающийся тем, что в качестве исходных защищенных олигонуклеотидов используют 5'-O-диметокситритил, 3'-Р (S)SMe, N-ацил, P(S)(SMe)-защищенные фосфодитиоатные олигонуклеотиды общей формулы III, где B' - остаток тимина, N4-бензоилцитозина, N6-бензоиладенина, N2-изобутирилгуанина; DMTr - 4,4'-диметокситритил; n = 1-20, а обработку раствором йода ведут в течение 1-2 ч. Технический результат - увеличение выхода целевого продукта и упрощение ...

СПОСОБ ПОЛУЧЕНИЯ S -(+)-2, 2-ДИМЕТИЛЦИКЛОПРОПАНКАРБОКСАМИДА, ФРАГМЕНТ ДНК, КОДИРУЮЩИЙ R -(-)-2,2-ДИМЕТИЛЦИКЛОПРОПАНКАРБОКСАМИДГИДРОКСИЛАЗУ COMAMONAS, РЕКОМБИНАНТНАЯ ПЛАЗМИДНАЯ ДНК (ВАРИАНТЫ), ШТАММ БАКТЕРИЙ ESCHERICHIA COLI (ВАРИАНТЫ)

Изобретение относится к генно-инженерному способу получения S-(+)2,2-диметилциклопропанкарбоксамида (S-(+)-2,2-ДМЦПКА). Последний является исходным продуктом для получения ингибитора дегидропептидазы циластатина, используемого в терапии для предотвращения дезактивации антибиотиков в почках под действием ренальной дегидропептидазы. В среду, содержащую рацемический R1S -(±)-2,2-ДМЦПКА, при рН 6-11 и температуре 15-55oС добавляется микроорганизм, трансформированный рекомбинантной плазмидной ДНК, содержащей фрагмент ДНК, кодирующий R-(-)-2,2-диметилциклопропанкарбоксамидгидролазу Comamonas, или экспрессируемую указанным микроорганизмом иммобилизованную гидролазу. Получают смесь S-(+)-2,2-ДМЦПКА и R-(-)-2, 2-диметилциклопропанкарбоновой кислоты, из которой выделяют конечный продукт. Способ осуществлен с помощью нового фрагмента ДНК, кодирующего ген гидролазы Comamonas, новых гибридных рекомбинантных плазмид, содержащих эту ДНК, новых микроорганизмов -штаммов Escherichia coli DSM N 7053 и E.coli ...

СПОСОБ ПОЛУЧЕНИЯ АДЕНОЗИН-5`-ТРИФОСФАТА

Изобретение относится к биотехнологии, а именно к способам получения аденозин-5'-трифосфата (АТФ) с помощью микроорганизмов. Цель изобретения - упрощение способа получения АТФ. Способ заключается в обработке водной суспензии дрожжевой культуры, выращенной на углеводоминеральной среде, озоновоздушной смесью с концентрацией озона 5· 108-5·109 молекул O3 на клетку так, чтобы активность дыхания дрожжей снизилась на 10 - 50%, после чего дрожжевые клетки удаляют, а АТФ выделяют из супернатанта известным способом. При этом не рекомендуется выдерживать водную суспензию дрожжей более 2 ч, так как снижается выход АТФ. Способ прост в осуществлении, так как сложная среда инкубации с дорогостоящими предшественниками АТФ-аденином или аденозином - заменяется дистиллированной водой, а инициатором синтеза АТФ является озон, к тому же накопление целевого продукта в воде упрощает его выделение и очистку. Способ является экологически чистым.

СИНТЕЗАТОР ОЛИГО/ПОЛИ/НУКЛЕОТИДОВ

Использование: в синтезе олиго (поли)нуклеотидов в молекулярной биологии, генной инженерии, медицине. Сущность: синтезатор олиго (поли)нуклеотидов, содержащий реактор 9 проточного типа, в рабочем объеме которого размещен твердофазный носитель, связанный с нуклеозидным компонентом, смеситель 6 для быстрого смешивания реагентов, содержащий камеру 7 на входе и пористую перегородку 8, размещенную на входе реактора 9, блок 1 размещения и подачи реагентов, блок 2 размещения и подачи промывающей жидкости, каналы 3 для подачи реагентов, промывающей жидкости и вывода отработанных реактивов, запорные клапаны 4 на каналах, блок 12 нагрева и поддержание заданной температуры реактивов, блок 11 управления. Синтезатор дополнительно содержит микродозатор 10, вход которого соединен с выходом реактора 9, а выход - с каналом 3 вывод отработанных реактивов, и гидростатические клапаны 5, причем вход каждого гидростатического клапана 5 соединен с каналом 3 подачи соответствующего реагента, а также каналом 3 ...

Способ очистки нуклеиновой кислоты

Изобретение относится к биотехнологии, а именно к композиции, содержащей высококонцентрированный ингибитор альфа-1 протеиназы (A1PI), и способу ее приготовления. Способ предусматривает использование однопроходной фильтрации в тангенциальном потоке. Изобретение эффективно для лечения состояний, связанных с лечением заболеваний, связанных с дефицитом A1PI, или заболеваний, при которых полезным является повышение уровня A1PI. 4 з.п. ф-лы, 2 ил., 6 табл., 5 пр.

СИНТЕТИЧЕСКИЕ ОЛИГОНУКЛЕОТИДНЫЕ ПРАЙМЕРЫ И СПОСОБ ВЫСОКОЧУВСТВИТЕЛЬНОГО ЭКСПРЕСС-ВЫЯВЛЕНИЯ ДНК ВИРУСА АФРИКАНСКОЙ ЧУМЫ СВИНЕЙ МЕТОДОМ ПЕТЛЕВОЙ ИЗОТЕРМИЧЕСКОЙ АМПЛИФИКАЦИИ В ПРИСУТСТВИИ ДНК ВНУТРЕННЕГО КОНТРОЛЬНОГО ОБРАЗЦА

Изобретение относится к области молекулярной диагностики. Описаны набор олигонуклеотидных праймеров и зонды для выявления ДНК вируса африканской чумы свиней (АЧС) и ДНК бактериофага лямбда как внутреннего контрольного образца исследования. Также предложен способ выявления ДНК вируса АЧС из биологического материала с использованием набора олигонуклеотидных праймеров и зондов с помощью петлевой изотермической амплификации (LAMP) в присутствии внутреннего контрольного образца ДНК бактериофага лямбда. Технический результат заключается в разработке высокочувствительного, специфичного и быстрого метода выявления вируса африканской чумы свиней. 2 н. и 1 з.п. ф-лы, 2 ил., 6 табл., 3 пр.

КОМПОЗИЦИЯ ДЛЯ ЛЕЧЕНИЯ СОСТОЯНИЙ, ЧУВСТВИТЕЛЬНЫХ ИЛИ ВОСПРИИМЧИВЫХ К ВОЗДЕЙСТВИЮ ЛОЖНОСПАРЕННОЙ DS РНК

Изобретение относится к медицине. Цель - повышение стабильности. Композиция содержит носитель, ложноспаренную dsРНК, поверхностно-активное вещество в определенных количествах. Ложноспаренная dsРНК содержит участки разрыва связи и имеет формулу rIn•r (C11-14• V)n, где n - целое число от 4 до 29. 2 табл.

ОЛИГОНУКЛЕОТИДЫ, ФАРМАЦЕВТИЧЕСКАЯ КОМПОЗИЦИЯ

Изобретение относится к олигонуклеотидам и их производным для подавления экспрессии изопренилпротеинтрансфераз, фармацевтическим композициям, содержащим такие соединения, и к использованию таких композиций олигонуклеотидов для лечения или терапии заболеваний человека или животных, вызванных ненормальной и/или нерегулируемой пролиферацией. Олигонуклеотиды имеют формулу (I): P - O-R (I), где Р либо отсутствует, либо представляет собой последовательность из одного или более оснований, выбранных из А (аденин), Т/(тимин), С (цитозин) или G (гуанин); О является последовательностью, выбранной из: SEQIDN1: AGAAGCCATG AT; SEQIDN2: GGACGTGACT GT; SEQIDN3: AAGGAGTAGC AG; SEQIDN4: CAGGCAGTAG CA; SEQIDN5: CCCGTCGTCC AT; SEQIDN6: CTGTCCCTGT AC; SEQIDN7: ACTTCTCTCT CC; SEQIDN8: ACTTGGTCCT AA; SEQIDN9: GCCATCATTC TG; SEQIDN10: GATCTGGACC AC; SEQIDN11: CTCAATAGCA TC; SEQIDN12: GTTTTTGGGG TG; SEQIDN13: TCTCACATCC TC; SEQIDN14: TTGTAAGAAC TG; SEQIDN15: GAAGTGCTTC TC; SEQIDN16: GCCTCTTTC AG; SEQIDN17: GGCATCTGTC ...

БИОЛОГИЧЕСКИ АКТИВНОЕ СОЕДИНЕНИЕ, СОДЕРЖАЩЕЕ КОДИРУЮЩИЙ ОЛИГОНУКЛЕОТИД (ВАРИАНТЫ), СПОСОБ ЕГО СИНТЕЗА, БИБЛИОТЕКА СОЕДИНЕНИЙ (ВАРИАНТЫ), СПОСОБ ЕЕ СИНТЕЗА, И СПОСОБ ПОИСКА СОЕДИНЕНИЯ, СВЯЗЫВАЮЩЕГОСЯ С БИОЛОГИЧЕСКОЙ МИШЕНЬЮ (ВАРИАНТЫ)

... 1. Способ синтеза соединения, содержащего функциональный компонент, который эффективно связан с кодирующим олигонуклеотидом, отличающийся тем, что в качестве исходного соединения используют соединение-инициатор, состоящее из исходного функционального компонента, содержащего n структурных фрагментов, где n равно целому числу 1 или более, по крайней мере одну реакционноспособную группу, и эффективно связанного с исходным олигонуклеотидом; осуществляют взаимодействие указанного соединения-инициатора со структурным фрагментом, содержащим по крайней мере одну реакционноспособную группу, комплементарную реакционноспособной группе соединения-инициатора, в условиях, пригодных для образования ковалентной связи между реакционноспособными группами; осуществляют взаимодействие исходного олигонуклеотида с новым вводимым олигонуклеотидом, который определяет структурный фрагмент, ковалентно связанный с соединением инициатором, причем взаимодействие исходного олигонуклеотида с новым олигонуклеотидом осуществляют ...

ОЛИГОНУКЛЕОТИДЫ, ТЕРАПЕВТИЧЕСКИЕ КОМПОЗИЦИИ, ПРИМЕНЕНИЕ КОМПОЗИЦИЙ

Изобретение относится к олигонуклеотидам и их производным для подавления экспрессии изопренил-протеин-трансфераз, терапевтическим композициям, содержащим такие соединения, и к использованию таких композиций олигонуклеотидов для лечения или терапии заболеваний человека или животных, вызванных ненормальной и/или нерегулируемой пролиферацией.

СПОСОБ ВВЕДЕНИЯ В ОЛИГОНУКЛЕОТИД АЛКИЛФОСФОНОТИОАТНОЙ ИЛИ АРИЛФОСФОНОТИОАТНОЙ МЕЖНУКЛЕОТИДНОЙ СВЯЗИ, СПОСОБ ПОЛУЧЕНИЯ ОЛИГОНУКЛЕОТИДА, ОЛИГОНУКЛЕОТИДЫ, СПОСОБ ИНГИБИРОВАНИЯ ГЕННОЙ ЭКСПРЕССИИ, СПОСОБ ЛЕЧЕНИЯ

В изобретении предложены усовершенствованные олигонуклеотиды, которые обладают повышенной устойчивостью к нуклеолитическому расщеплению за счет наличия алкилфосфонотиоатных или арилфосфонотиоатных межнуклеотидных связей.

НУКЛЕИНОВЫЕ КИСЛОТЫ, МОДИФИЦИРОВАННЫЕ АМИНОКИСЛОТАМИ

... 1. Соединение формулы I в любой из групп I-V: группа I, где X обозначает CHR2OH, где R2 обозначает H, (низший алкил)амин или (низший алкил)имидазол; Y обозначает Base-(CHR1)n-, где Base обозначает негалогенезированное вариабельное нуклеозидное основание, R1 обозначает Н, низший алкил, (низший алкил)амин или (низший алкил)имидазол, n равно 1-7 С атомов; А обозначает карбонил; Z обозначает Н или OR3, где R3 обозначает Н, низший алкил, (низший алкил)амин или (низший алкил)имидазол; W обозначает CHR4OH, где R4 обозначает Н, (низший алкил)амин или (низший алкил)имидазол; N обозначает N (азот); и L ничего не обозначает; группа II, где X обозначает Base-(CHR1)n-, где Base обозначает негалогенезированное вариабельное нуклеозидное основание, R1 обозначает Н, низший алкил, (низший алкил)амин или (низший алкил)имидазол, n равно 1-7 С атомов; Y обозначает CHR2OH, где R2 обозначает H, (низший алкил)амин или (низший алкил)имидазол; А обозначает карбонил или CH2; Z обозначает Н или OR3, где R3 обозначает ...

НУКЛЕОТИДНАЯ ПОСЛЕДОВАТЕЛЬНОСТЬ ДЛЯ КОНТРОЛЯ ЭКСПРЕССИИ

... 1. Нуклеотидная последовательность для контроля экспрессии, которая осуществляет контроль экспрессии целевого гена, расположенного после последовательности для контроля экспрессии, зависящей от внутриклеточной концентрации аминокислоты, причем в бактерии, содержащей ДНК-конструкцию с нуклеотидной последовательностью для контроля экспрессии, промотором расположенным перед нуклеотидной последовательностью для контроля экспрессии, и целевым геном, расположенным после нуклеотидной последовательности для контроля экспрессии, частота терминации на нуклеотидной последовательности для контроля экспрессии, при транскрипции, начинающейся на указанном промоторе, понижается при увеличении внутриклеточной концентрации аминокислоты, посредством чего экспрессия целевого гена повышается. 2. Нуклеотидная последовательность для контроля экспрессии по п. 1, отличающаяся тем, что указанная последовательность содержит область, кодирующую лидерный пептид, содержащий указанную аминокислоту и ρ-независимый терминатор ...

Носитель для твердофазного синтеза, способ его получения и его применение

Изобретение относится к области получения функциональных полимерных материалов, а именно, представляет собой носитель для использования в твердофазном синтезе олигонуклеотидов. Носитель для твердофазного синтеза описывается формулой: где R1= -(CH2)n-, n – целое число о 0 до 3, или R1= -O-(CH2)m-O-, m – целое число от 1 до 4; и R2= -OH или -NH2; при этом получение носителя для твердофазного синтеза включает следующие этапы: вначале получают водную и масляную фазы; водная фаза содержит воду, диспергирующее средство и неорганическую соль; масляная фаза содержит: поперечно-сшивающий мономер, моновинильное соединение, функциональный мономер, модифицированный мономер; порообразующий агент и инициатор. Затем масляную фазу добавляют к водной при перемешивании и нагревании для проведения реакции полимеризации, после чего, удаляют порообразующий агент. В результате получают пористую полимерную смолу, которая способна подвергаться дальнейшей реакции для получения носителя твердофазного синтеза, содержащего ...

ОЛИГОДЕЗОКСИРИБОНУКЛЕОТИДНЫЙ ИНГИБИТОР ДНК-МЕТИЛТРАНСФЕРАЗЫ 1 ЧЕЛОВЕКА

Изобретение относится к области молекулярной биологии и может быть использовано в медицине. Предложен олигодезоксирибонуклеотидный ингибитор ДНК-метилтрансферазы 1 человека (Dnmt1), характеризующийся тем, что он имеет самокомплементарную структуру, которая образует двуцепочечную шпильку, имеет неспаренную СА пару в сайте узнавания фермента ДНК-метилтрансферазы 1 человека и содержит в своем составе 5-метилцитозин (5mC) и тиофосфаты вместо фосфатов. Изобретение обеспечивает селективное ингибирование активности фермента Dnmt1 в реакцияхи. 3 ил., 3 пр.

Химическое соединение с триазиновой группой и способ его получения

ИММУНОЛОГИЧЕСКИЕ АНТИГЕНЫ ВИРУСА ПРОСТОГО ГЕРПЕСА И СПОСОБЫ ИХ ИСПОЛЬЗОВАНИЯ

... 1. Фармацевтическая композиция, включающая выделенный полипептид вируса простого герпеса (HSV), где полипептид включает белок UL19, UL21, UL49 или UL50 или его фрагмент и фармацевтически приемлемый носитель. 2. Фармацевтическая композиция, включающая выделенный полипептид HSV и фармацевтически приемлемый носитель, где полипептид включает аминокислотную последовательность, выбранную из группы, состоящей из: (а) аминокислоты 1078-1319 UL19;(б) аминокислоты 148-181 UL21;(в) аминокислоты 105-190 или 177-220 UL49;(г) аминокислоты 118-312 UL50;(д) аминокислоты 1-273 гликопротеина Е (gE);(е) аминокислоты 185-197, 209-221 или 430-449 VP16; и (ж) измененных вариантов (а)-(е). 3. Композиция по п.1 или 2, где полипептид является слитым белком. 4. Композиция по п.3, где слитый белок является растворимым. 5. Полинуклеотид, который кодирует полипептид, включающий аминокислотную последовательность, выбранную из группы, состоящей из: (а) аминокислоты 1078-1319 UL19;(б) аминокислоты 148-181 UL21;(в) аминокислоты ...

СПОСОБ СИНТЕЗА ПЦР-ЗОНДОВ ПО РЕАКЦИИ [3+2]-ДИПОЛЯРНОГО ЦИКЛОПРИСОЕДИНЕНИЯ

Способ синтеза зондов для полимеразной цепной реакции режима реального времени, включающий медь-катализируемую реакцию [3+2]-диполярного циклоприсоединения между алкином и азидом, протекающую в инертной атмосфере в присутствии лигандов, стабилизирующих степень окисления меди(I).

АНАЛОГИ ОЛИГОНУКЛЕОТИДОВ КАК СРЕДСТВА КОРРЕКЦИИ СПЛАЙСИНГА ДЛЯ ЛЕЧЕНИЯ МЫШЕЧНОЙ ДИСТРОФИИ ДЮШЕНА

СПОСОБ СИНТЕЗА МОДИФИЦИРОВАННЫХ ПО АТОМУ ФОСФОРА НУКЛЕИНОВЫХ КИСЛОТ

... 1. Способ синтеза нуклеиновой кислоты, содержащей хиральную Х-фосфонатную составляющую, который включает реакцию молекулы, содержащей ахиральную H-фосфонатную составляющую, и нуклеозида, содержащего 5'-ОН составляющую, с получением конденсированного промежуточного соединения; и превращение данного промежуточного соединения в нуклеиновую кислоту, содержащую хиральпую Х-фосфонатную составляющую. 2. Способ по п.1, отличающийся тем, что стадия реакции молекулы, содержащей ахиральную H-фосфонатную составляющую, и нуклеозида, содержащего 5'-ОН составляющую, с получением конденсированного промежуточного соединения, представляет собой реакцию "в одном котле". 3. Способ по п.1, отличающийся тем, что нуклеиновая кислота, содержащая хиральную X-фосфонатную составляющую, является соединением формулы 1: где R1 представляет собой -ОН, -SH, -NRdRd, -N3, галоген, водород, алкил, алкенил, алкинил, алкил-Y1-, алкенил-Y1-, алкинил-Y1-, арил-Y1-, гетероарил-Y1-, -P(O)(Re)2, -HP(O)(Re), -ORa или -SRc; Y1 является ...

КОМПОЗИЦИИ И СПОСОБЫ

КОНЪЮГАТ ПОЛИПЕПТИД-НУКЛЕИНОВАЯ КИСЛОТА ДЛЯ ИММУНОПРОФИЛАКТИКИ ИЛИ ИММУНОТЕРАПИИ ДЛЯ НЕОПЛАСТИЧЕСКИХ ИЛИ ИНФЕКЦИОННЫХ НАРУШЕНИЙ

... 1. Композиция, содержащая: ! (а) нацеливающую часть, где указанная нацеливающая часть связывается с компонентом опухолевой клетки, микроокружения опухоли, сосудистой сети опухоли, опухолевым антигеном, связанным с опухолью антигеном, молекулой поверхности опухолевой клетки, опухолевой антигенной детерминантой или содержащим опухолевый антиген слитым белком; и ! (b) биологически активный агент, где указанный биологически активный агент содержит: ! i) полипептид антигена или антигенной детерминанты, где указанный полипептид антигена или антигенной детерминанты содержит компонент патогена, микроорганизма, антигена патогена, антигенной детерминанты патогена, связанного с патогеном антигена или содержащего антиген патогена слитого белка, ! и указанный полипептид антигена или антигенной детерминанты получен из вакцины; или ! ii) нуклеиновую кислоту, кодирующую полипептид антигена или антигенной детерминанты, ! где указанный полипептид антигена или антигенной детерминанты содержит компонент патогена ...

СПОСОБ ПОЛУЧЕНИЯ БИОТИНМЕЧЕННЫХ ОЛИГОНУКЛЕОТИДОВ

Изобретение относится к области биотехнологии, молекулярной биологии, медицинской диагностики и может быть использовано для идентификации исследуемого биологического материала, в частности при гибридизационном анализе нуклеиновых кислот, в том числе вирусных, с использованием детекторной авидин-биотиновой (стрептавидин-биотиновой) системы. Предложен способ получения биотинмеченных олигонуклеотидов путем взаимодействия биотинсодержащего реагента с аминогруппой спейсера, предварительно введенного в олигонуклеотид, при этом в качестве биотинсодержащего реагента используют 4-сульфо-3-нитрофениловый или сульфотетрафторфениловый эфир биотина, а процесс ведут в водном растворе. Данный способ обеспечивает возможность получения биотинмеченных олигонуклеотидов в препаративных количествах с высоким выходом и сокращением времени проведения процесса.

Способ получения фракций нуклеиновых кислот

Способ получения синтетической двунитчатой РНК, обладающей интерферониндуцирующей активностью

Способ получения фрагмента нуклеиновой кислоты, кодирующего проинсулин человека

Изобретение относится к генетической инженерии и касается получения фрагмента нуклеиновой кислоты, кодирующего проинсулин человека с помощью рекомбинантной ДНК-технологии. Фрагмент ДНК, кодирующий проинсулин, получают путем клонирования гена млекопитающего. После клонирования в плазмиде PBR 322 его выделяют с помощью эндонуклеаз HHA 1 и ALU 1 или эндонуклеаз HHA 1 и модифицируют, присоединяя химически синтезированную нуклеотидную последовательность 3Ъ-TTTGTGAACCAACACCTGTCGGCT CACACCTGGAAG-5Ъ, кодирующую 14 аминокислот проинсулина.

XANTHIN DERIVATE VERBINDUNGEN ZUR AKTIVIERUNG DER CHLORID-LEITUNG IN TIERZELLEN

XANTHIN DERIVATE VERBINDUNGEN ZUR AKTIVIERUNG DER CHLORID-LEITUNG IN TIERZELLEN

VERFAHREN ZUR KONTROLLIERTEN SYNTHESE VON BELIEBIGEN PEPTIDMISCHUNGEN KODIERENDE POLYNUKLEOTIDMISCHUNGEN

Vorrichtung, Verfahren und Einrichtung zur Isolierung von Nukleinsäuren

Vaskulärer endothelialer Wachstumsfaktor 2

ETIKETTIERREAGENZ UND NACHWEISMETHODE

7-DEAZAPURIN MODIFIZIERTE OLIGONUKLEOTIDE

VERFAHREN ZUR SCHNELLEN SYNTHESE VON GROSSEN ZAHLEN TRAEGERGEBUNDENER ODER FREIER PEPTIDE

The invention concerns a method for the rapid synthesis of substrate-bound or free peptides, apparatus for carrying out the method, a flat material with peptides prepared by the method bound to it, and a use of this material.

DNS-SEQUENZBESTIMMUNG DURCH MASSENSPEKTROMETRIE AUF DEM WEG DES ABBAUS MIT EXONUKLEASE

POLYNUKLEOTID REAGENZIEN MIT NICHTNUKLEOTIDISCHEN GRUPPIERUNGEN, DEREN HERSTELLUNG UND VERWENDUNG

OLIGO-2'-DESOXYNUKLEOTIDE UND IHRE VERWENDUNG ALS ARZNEIMITTEL MIT ANTIVIRALER WIRKSAMKEIT

The invention concerns oligodeoxyribonucleotides, in which, at both the 5' and the 3' terminal positions, at least two 2'-deoxy- beta -D-erythro-pentofuranosyl groups have been replaced by 2'-deoxy- beta -D-threo-pentofuranosyl groups, and oligodeoxyribonucleotides in which at least 20 % of the 2'-deoxy- beta -D-erythro-pentofuranosyl groups in successive nucleotide units have been replaced by 2'-deoxy- beta -D-threo-pentofuranosyl groups, and which consist of 6 to 100 nucleotide units. Such oligodeoxyribonucleotides are suitable for use as anti-sense inhibitors of the expression of viral genes and oncogenes and can be used in the preparation of drugs with an anti-viral action.

Markierte Oligonukleotide

VERFAHREN, TESTSÄTZE UND REAKTIVE TRÄGER ZUR 3'-MARKIERUNG VON OLIGONUKLEOTIDEN

2'-5'-Phosphorothioat-Oligoadenylate und deren antivirale Verwendungen

OLIGONUCLEOTID-POLYAMID KONJUGATE

DEM MENSCHLICHEN GEWEBEFAKTOR ÄHNLICHE DNS-SEGMENTE, POLYPEPTIDE UND ANTIKÖRPER

IN DER HAUPTKETTE MODIFIZIERTE OLIGONUKLEOTID-ANALOGE

ANTISENSE OLIGONUKLEOTIDE

DREI POLYMORPHE DNS-MARKER MIT REPETITIVEN MIKROSATELLITENSEQUENZEN MIT HOHER INFORMATIONSDICHTE

2-AZAPURINE VERBINDUNGEN UND IHRE VERWENDUNG

Auf einer Oberfläche befestigte multifunktionelle Polymere-Netzwerke für Sensorchips

Hydrophobe Nukleinsäure-Sonde

Verfahren zur Überprüfung der Kontamination von oligonukleotidabhängigen Nukleinsäure-Amplifikationsreaktionen

NUKLEOSID-DERIVATE UND IHRE VERWENDUNG IN DER OLIGONUKLEOSID-SYNTHESE.

OLIGONUKLEOTIDE SYNTHESE

Verfahren zur Herstellung von N-Glykosiden

Synthetisierungsvorrichtung in fester Phase

Effektive Gensonden-Konjugationen mittels eines aussergewöhnlichen Mischanhydridverfahrens

METHOD OF SYNTHESIZING WITHIN A BACTERIAL HOST A SELECTED MATURE PROTEIN OR POLYPEPTIDE

NANOPARTIKEL-POLYANION-KONJUGATE SOWIE VERFAHREN ZUR VERWENDUNG DAVON BEIM NACHWEIS VON ANALYTEN

0,0'-Dicyanoethyl-phosphoramidite, Verfahren zu deren Herstellung und ihre Verwendung in Phosphorylierungsreaktionen.

Einschrittverfahren und Polynukleotidverbindungen zur Hybridisierung mit Ziel-Polynukleotiden

Markierte Nukleinsäuren.

VERSTAERKUNG VON HYBRIDISIERUNGSSIGNALEN DURCH VERWENDUNG VON KOMPLEMENTARISCHEN DNS-STRAENGEN.

2'-MODIFIZIERTE NUKLEOTIDE

Sonden

Nachweis von Nukleinsäuren durch Fluoreszenz quenching

Nachweis von Nukleinsäuren durch Aufhebung der Fluoreszenz

Nachweis von Nukleinsäuren durch Aufhebung der Fluoreszenz

DEVICE FOR SIMULTANEOUSLY CARRYING OUT A PLURALITY OF CHEMICAL REACTION STEPS

Vorrichtung zum Nachweis von fluoreszenten, der Bindung eines Liganden auf einem Substrat entsprechenden Merkmalen

Therapeutische Anwendungen von 2',5'-Oligoadenylat-Derivaten

UNGELADENE, AUF MORPHOLIN BASIERENDE POLYMERE MIT CHIRALEN, PHOSPHOR ENTHALTENDEN BRÜCKEN ZWISCHEN DEN UNTEREINHEITEN

OLIGONUKLEOTID-ANALOGE.

PLATINENTHALTENDE VERBINDUNG, VERFAHREN ZU DEREN HERSTELLUNG SOWIE DEREN VERWENDUNG

ANTISENSE-MODULATION DER SURVIVIN-EXPRESSION

New 2-(1-((hetero)aryl(oxy))-ethyl)-nitrobenzene derivatives, useful for introducing intramolecular triplet sensitized photolabile protecting groups into synthons for light-controlled biopolymer production

... 2-(1-((Hetero)aryl or (hetero)aryloxy)-ethyl)-nitrobenzene derivatives (I) are new. Also new are nucleotide synthon compounds (II) containing a photolabile protecting group derived from (I). Nitrobenzene derivatives of formula (I) are new. R1 = halo, CN or NO2; or alkyl, alkenyl, alkynyl, alkoxy, alkoxycarbonyl, aryl, aryloxy, heteroaryl or heteroaryloxy (all optionally substituted, aryl or heteroaryl moieties being mono- or polycyclic); or adjacent groups R1 may together form a 5- or 6-membered ring (optionally containing heteroatoms); R2 = mono- or polycyclic aryloxy or heteroaryloxy; or polycyclic aryl or heteroaryl containing at least 3 fused rings; R3 = trialkylsilyl; or alkyl, alkenyl, alkynyl, alkoxy, alkoxycarbonyl, aryl, aryloxy, heteroaryl or heteroaryloxy (all optionally substituted, aryl or heteroaryl moieties being mono- or polycyclic); m = 0-4; p = 0 or 1; X = -C(O)-, -O-C(O)-, -O-C(S)- or -S(O)2-; and Y = H, leaving group or substrate. An Independent claim is included for ...

Rekombinante Impfstoffe und deren Verwendung

Fusionsmolekül, das ein Antigen, eine Transmembranregion und die cytoplasmatische Region einer Kette eines MHC-Moleküls umfasst.

New 3'-derivatised oligo:nucleotide analogues

... 3'-Modified oligonucleotides (I) and (II) and their salts are new. In the formula, Q = gp. (a), i.e. cpds. (I), or gp. (b), i.e. cpds. (II); a,b = 0-20; R<1> = H; alkyl, alkenyl or alkynyl with up to 18C; 1-18C alkylcarbonyl; 2-19C alkenylcarbonyl; 3-19C alkynylcarbonyl; 6-20C aryl; 6-14C aryl-1-8C alkyl; or a gp. -P(W)(Z)(Z') (III); R<2> = H; OH; 1-18C alkoxy; halo; N-3; or NH2; D = OH or OPO3<2->; B = conventional nucleotide base linked in the alpha- or beta-configuration; n = 1-100; n' = 0-50; m = 0-5; m' = 0-5 in cpds. (I) and 1-5 in cpds. (II); A = O; S or CH2; W = O, S or Se; V = O or sulphanediyl; T = O; sulphanediyl; NH; or CH2; X = OH or SH; U = OH; SH; BH3; SeH; alkyl or alkoxy with up to 18C; 6-20C aryl; 6-14C aryl-1-8C alkyl; NHR<3>; NR<3>R<4>; or a gp. (OCH2CH2)pO(CH2)qCH2R<5> (IV); R3 = 1-18C alkyl; 6-20C aryl; 6-14C aryl-1-8C alkyl; or (CH2)c-NH(CH2)cd-NR<6>R<6>; and R<4> = 1-18C alkyl; 6-20C aryl; or 6-10C aryl-1-8C alkyl; or NR<3>R<4> forms a 5- or 6-membered heterocycle ...

NEUE LOW DENSITY LIPOPROTEIN BINDENDE PROTEINE UND DEREN VERWENDUNG IN DER DIAGNOSTIK UND BEHANDLUNG VON ATHEROSKLEROSE

ANTIBAKTERIELLE PROTONIERTE OLIGONUKLEOTIDE

SULFONATFREIE CYANINFARBSTOFFE ZUR MARKIERUNG VON NUKLEOSIDEN UND NUKLEOTIDEN

NEUE PSORALENDERIVATE

Massives Parallelverfahren zur Dekodierung von DNA und RNA

Ein Nukleotidanalogon, welches umfasst: a) eine Base ausgewählt aus der Gruppe bestehend aus Adenin oder einem Analogon von Adenin, Cytosin oder einem Analogon von Cytosin, Guanin oder einem Analogon von Guanin, Thymin oder einem Analogon von Thymin sowie Uracil oder einem Analogon von Uracil; b) eine eindeutige Markierung, welche mittels eines abspaltbaren Linkers an der Base oder an einem Analogon der Base befestigt ist; c) eine Desoxyribose; und d) eine abspaltbare chemische Gruppe, um eine -OH-Gruppe an einer 3'-Position der Desoxyribose zu schützen, wobei die abspaltbare chemische Gruppe durch chemische Mittel abgespalten werden kann.

Methods of creating and screening dna-encoded libraries

The present invention features a number of methods for identifying one or more compounds that bind to a biological target. The methods include synthesizing a library of compounds, wherein the compounds contain a functional moiety having one or more diversity positions. The functional moiety of the compounds is operatively linked to an initiator oligonucleotide that identifies the structure of the functional moiety.

Methods of using oligomeric compounds comprising 2'-substituted nucleosides

The present disclosure provides oligomeric compounds comprising at least one 2′-fluoroethoxy modified nucleoside of formula I and methods of using these oligomeric compounds. The methods provided herein include contacting a cell or administering to an animal at least one of the oligomeric compounds. In certain embodiments, the oligomeric compounds hybridize to a portion of a target RNA resulting in loss of normal function of the target RNA.

Flow Chart Programming Platform for Testers and Simulators

A system for the development, compilation, execution, monitoring and debug of automated test and simulation systems in a flow chart programming language. A development and debug system, implemented as software on a computer, which provides an application developer the capability to enter fully defined application programs through the use of graphical flow charts. An executions system, implemented as a program on a device incorporating a central processing unit, memory, communications and necessary interfaces, which executes graphical flow charts compiled by the development and debug system. The development and debug system communicates with the execution system to download programs, control operation, monitor operation and provide a debugging environment.

Preparation and isolation of 5' capped mrna

The synthesis of capped/tagged RNA, methods of use and kits providing same are contemplated. Tagged RNA permits isolation of RNA transcripts in vitro. The ability to isolate and purify capped RNA results in improved transcription and translation and provides a tool for identifying RNA-protein interactions. Such capped RNA finds use in therapeutic applications, diagnosis and prognosis and in the treatment of cancers and HIV.

Modulation of factor 7 expression

Disclosed herein are antisense compounds and methods for decreasing Factor 7 and treating or preventing thromboembolic complications in an individual in need thereof. Examples of disease conditions that can be ameliorated with the administration of antisense compounds targeted to Factor 7 include thrombosis, embolism, and thromboembolism, such as, deep vein thrombosis, pulmonary embolism, myocardial infarction, and stroke. Antisense compounds targeting Factor 7 can also be used as a prophylactic treatment to prevent individuals at risk for thrombosis and embolism.

Synthesis of oligonucleotides

A method for preparing an oligonucleotide comprising the steps of a) providing a hydroxyl containing compound having the formula (A), b) reacting said compound with a phosphitylating agent in the presence of an activator (activator I) having the formula (I) to prepare a phosphitylated compound; and c) reacting said phosphitylated compound without isolation with a second compound having the formula (A) wherein R 5 , R 3 , R 2 , B are independently selected, but have the same definition as above in the presence of an activator II different from activator I.

In vivo gene regulation by the combination of knock-in-teto sequence into the genome and tetracycline-controlled trans-suppressor (tts) protein

Disclosed is a FAST (Flexible Accelerated STOP TetO-knockin) system, an efficient method for manipulating gene expression in vivo to rapidly screen animal models of disease. This invention further discloses a single gene targeting event yielding 2 distinct knockin mice—STOP-tetO and tetO knockin—which permit generation of multiple strains with variable expression patterns: 1) knockout, 2) Cre-mediated rescue; 3) tTA-mediated misexpression; 4) tTA-mediated overexpression; and 5) tTS-mediated conditional knockout/knockdown. Using the FAST system, multiple gain- and loss-of-function strains can therefore be generated on a timescale not previously achievable. These strains can then be screened for clinically-relevant abnormalities. The flexibility and broad applicability of the FAST system is demonstrated by targeting several genes encoding proteins implicated in neuropsychiatric disorders: Mlc1, Neuroligin 3, the serotonin 1A receptor, and the serotonin 1B receptor.

MODULATION OF INSULIN LIKE GROWTH FACTOR I RECEPTOR EXPRESSION

The present invention provides compositions and methods for modulating the expression of growth factor gene. In particular, this invention relates to compounds, particularly oligonucleotide compounds, which, in preferred embodiments, hybridize with nucleic acid molecules encoding the Insulin Like Growth Factor I receptor (IGF-I receptor or IGF-IR) and in particular human IGF-IR. Such compounds are exemplified herein to modulate proliferation which is relevant to the treatment of proliferative and inflammatory skin disorders and cancer. It will be understood, however, that the compounds can be used for any other condition in which the IGF-IR is involved including inflammatory condition. 144-. (canceled)45. An antisense oligonucleotide comprising 20 to 80 nucleobases in length targeted to a nucleic acid molecule encoding human insulin-like growth factor receptor (IGF-IR) , wherein said oligonucleotide specifically hybridizes with said nucleic acid molecule and inhibits the expression of IGF-IR and wherein said oligonucleotide comprises a deoxynucleotide region flanked on both the 5′ and the 3′ ends of said deoxynucleotide region with 2′-O-(2-methoxyethyl) nucleotides.46. The oligonucleotide as claimed in claim 45 , wherein the oligonucleotide is 20 to 30 nucleobases in length.47. The oligonucleotide as claimed in claim 45 , wherein the oligonucleotide is 8 to 30 nucleobases in length.48. The oligonucleotide as claimed in claim 45 , wherein said oligonucleotide comprises a ten deoxynucleotide region flanked on both the 5′ and the 3′ ends of said ten deoxynucleotide region with five 2′-O-(2-methoxyethyl) nucleotides.49. The oligonucleotide of which is a DNA oligonucleotide claim 45 , an RNA oligonucleotide or a chimeric oligonucleotide.50. The oligonucleotide of claim 45 , wherein the nucleic acid molecule is an RNA molecule encoding human insulin-like growth factor receptor.51. The oligonucleotide of claim 50 , wherein at least a portion of said oligonucleotide ...

Particle matrix for storage of biomolecules

Matrices for manipulation of biopolymers, including the separation, purification, immobilization and archival storage of biopolymers is disclosed.

Procedure for the specific isolation of total dna content of bacterial germs and a kit for this purpose

Procedure for the specific isolation of total DNA content of bacterial germs of different samples, in the course of which the cells are lysated, the DNA content of the lysate is bound selectively, it is washed and then the desalinated linear polymer nucleic acid is eluted from the binding surface in an aqueous solution. Before cell lysis the nonviable bacterial cells are separated from the viable cells on the basis of their different cell surface physical-chemical characteristics, the viable cells of the sample are kept and then lysated using a mechanical and/or enzymatic, favourably lysozyme enzymatic method. After this exclusively double-stranded DNA deriving from the lysate of viable cells is bound on a —SiO 2 —TiO 2- matrix containing chemically activated —OH and dodecylamine groups, and after washing it, the desalinated linear polymer nucleic acid is eluted in an aqueous solution.

5' MODIFIED NUCLEOSIDES AND OLIGOMERIC COMPOUNDS PREPARED THEREFROM

The present invention provides 5′ modified nucleosides and oligomeric compounds prepared therefrom. More particularly, the present invention provides modified nucleosides having at least one 5′-substituent and an optional 2′ substituent, oligomeric compounds comprising at least one of these modified nucleosides and methods of using the oligomeric compounds. In some embodiments, the oligomeric compounds provided herein are expected to hybridize to a portion of a target RNA resulting in loss of normal function of the target RNA. 146-. (canceled)48. The 5′ modified nucleoside of wherein Bx is uracil claim 47 , thymine claim 47 , cytosine claim 47 , 5-methylcytosine claim 47 , adenine or guanine.49. The 5′ modified nucleoside of wherein qand qare each claim 48 , independently claim 48 , OCH claim 48 , OCHCHor OC(H)(CH)and qis O.50. The 5′ modified nucleoside of wherein r is 1 claim 49 , Mis H and Mis hydroxyl or r is 0 claim 49 , Mis O(CH)CN and Mis N[CH(CH)].51. The 5′ modified nucleoside of wherein G is halogen claim 50 , OCH claim 50 , OCHF claim 50 , OCHF claim 50 , OCF claim 50 , OCHCH claim 50 , O(CH)F claim 50 , OCHCHF claim 50 , OCHCF claim 50 , OCH—CH═CH claim 50 , O(CH)—OCH claim 50 , O(CH)—SCH claim 50 , O(CH)—OCF claim 50 , O(CH)—N(R)(R) claim 50 , O(CH)—ON(R)(R) claim 50 , O(CH)—O(CH)—N(R)(R) claim 50 , OCHC(═O)—N(R)(R) claim 50 , OCHC(═O)—N(R)—(CH)—N(R)(R) or O(CH)—N(R)—C(═NR)[N(R)(R)] wherein R claim 50 , R claim 50 , Rand Rare each claim 50 , independently claim 50 , H or C-Calkyl.52. The 5′ modified nucleoside of wherein G is F claim 51 , OCH claim 51 , O(CH)—OCH claim 51 , OCHC(═O)—N(H)CHor OCHC(═O)—N(H)—(CH)—N(CH).53. The 5′ modified nucleoside of wherein g is 1.54. The 5′ modified nucleoside of wherein three of q claim 53 , q claim 53 , qand qare each H.55. The 5′ modified nucleoside of wherein one of qand qis H claim 53 , one of qand qis H and the other two of q claim 53 , q claim 53 , qand qare other than H.56. The 5′ modified nucleoside of wherein ...

Photogenerated reagents

This invention describes reagent precursors and methods for chemical and biochemical reactions. These reagent precursors that can be activated in solution upon irradiation to generate reagents required for the subsequent chemical reactions. Specifically, photogenerated reagents (PGR) are useful for controlling parallel combinatorial synthesis and various chemical and biochemical reactions. 164-. (canceled)65. A method for forming a nucleic acid polymer comprising:(a) derivatizing a solid surface with a protected moiety containing at least one protective group;(b) contacting the solid surface with solution comprising at least one photogenerated reagent precursor and at least one co-reagent;(c) irradiating the solution thereby generating a photogenerated reagent in at least a portion of the solution such that the protective group is removed from the protected moiety; and(d) coupling a nucleic acid monomer to the deprotected moiety.661. The method of claim wherein the photogenerated reagent precursor is a photogenerated reagent acid precursor.671. The method of claim wherein the co-reagent is a base.681. The method of claim wherein the protected moiety is a protected nucleotide.691. The method of claim wherein the photogenerated reagent precursor is a photogenerated reagent base precursor.701. The method of claim wherein the co-reagent is an acid.711. The method of claim wherein the protected moiety is a protected nucleic acid.721. The method of claim wherein the co-reagent is a sensitizer or supersensitizer.73. A method of deprotecting protected nucleic acids attached to a solid surface and wherein the nucleic acids have one or more protecting group(s) comprising a) contacting the protecting group moieties with a solution comprising one or more PGR-P(s) and one or more co-reagent(s) and b) irradiating the solution to produce one or more PGR(s) wherein the PGR(s) remove the protecting group(s) from the protected nucleic acids.749. The method of claim wherein the ...

SUBSTITUTED 2'-AMINO AND 2'-THIO-BICYCLIC NUCLEOSIDES AND OLIGOMERIC COMPOUNDS PREPARED THEREFROM

Provided herein are 2′-amino and 2′-thio bicyclic nucleosides and oligomenc compounds prepared therefrom. The novel bicyclic nucleosides provided herein are expected to be useful for enhancing one or more properties of the oligomeric compounds they are incorporated into such as nuclease resistance. 192-. (canceled)94. The bicyclic nucleoside of wherein Bx is an optionally protected uracil claim 93 , thymine claim 93 , cytosine claim 93 , 5-methylcytosine claim 93 , adenine or guanine.95. The bicyclic nucleoside of wherein Tis 4 claim 93 ,4′-dimethoxytrityl and Tis diisopropylcyanoethoxy phosphoramidite.96. The bicyclic nucleoside of wherein Qand Qare each H.97. The bicyclic nucleoside of wherein one of Qand Qis H and the other of Qand Qis C-Calkyl or substituted C-Calkyl.98. The bicyclic nucleoside of wherein one of Qand Qis CH.99. The bicyclic nucleoside of wherein Gand Gare each H.100. The bicyclic nucleoside of wherein one of Gand Gis H and the other of Gand Gis C-Calkyl or substituted C-Calky.101. The bicyclic nucleoside of wherein at least one of Gand Gis CH.102. The bicycle nucleoside of wherein Z is NR wherein R is H claim 93 , C-Csubstituted C-Calkyl claim 93 , C-Calkoxy or substituted C-Calkoxy.103. The bicyclic nucleoside of wherein R is CHor methoxy.104. The bicyclic nucleoside of wherein Z is S.106. The bicyclic nucleoside of wherein three of Q claim 105 , Q claim 105 , Gand Gare H and the other one of Q claim 105 , Q claim 105 , Gand Gis CH.107. The bicyclic nucleoside of wherein two of Q claim 105 , Q claim 105 , Gand Gare H and the remaining two of Q claim 105 , Q claim 105 , Gand Gare CHwherein the two that are CHare selected from Qand G claim 105 , Qand G claim 105 , Qand G claim 105 , and Qand G.109. The oligomeric compound of wherein Bx is an optionally protected uracil claim 108 , thymine claim 108 , cytosine claim 108 , 5-methylcytosine claim 108 , adenine or guanine for each bicyclic nucleoside of Formula II.110. The oligomeric compound of ...

Rapid nucleic acid purification

Provided is a method for rapid nucleic acid purification, and the method for rapid nucleic acid isolation according to the present invention is very useful in diagnosing causes of disease or detecting a target gene; can be used in molecular diagnosis of causes of disease more rapidly and conveniently, as compared with the existing nucleic acid isolation method requiring complicated and special equipment; does not require skills therefor, thereby allowing an ordinary person to personally conduct isolation of nucleic acid for analyzing causes of disease and further solving the existing inconvenience caused by directly going to the hospitals or health clinical centers; and can analyze causes of disease more promptly.

Methods, Reagents and Kits for Preservation of Nucleic Acids in Biological Samples

Provided is a nucleic acid preservative comprising at least one reducing agent, at least one chaotropic substance, at least one polyamine substance and at least one chelating agent and uses thereof, and a method for the preservation of nucleic acids in a biological sample. Further provided are kits for use in the preservation of nucleic acids in a biological sample, and more particularly, a blood sample. 1. A nucleic acid preservative comprising at least one reducing agent , at least one chaotropic substance , at least one polyamine substance and at least one chelating agent.2. The nucleic acid preservative according to claim 1 , wherein the reducing agent is glutathione and wherein the glutathione is present in an amount from about 10 mM to about 200 mM.3. The nucleic acid preservative according to claim 1 , wherein the chaotropic substance is a lithium salt and wherein the lithium salt is present in an amount from about 1 M to about 4 M.4. The nucleic acid preservative according to claim 3 , wherein the lithium salt is LiCl.5. The nucleic acid preservative according to claim 1 , wherein the chaotropic substance is a guanidium salt claim 1 , and wherein the guanidium salt is present in an amount from about 0.1 M to about 0.9 M.6. The nucleic acid preservative according to claim 5 , wherein the guanidium salt is guanidine hydrochloride.7. The nucleic acid preservative according to claim 1 , wherein the chaotropic substance is urea and wherein the urea is present in an amount from about 2 M to about 12 M.8. The nucleic acid preservative according to claim 1 , wherein the polyamine substance is spermidine and wherein the spermidine is present in an amount from about 10 μM to about 300 μM.9. The nucleic acid preservative according to claim 1 , wherein the polyamine substance is biuret and wherein the biuret is present in an amount of about 10 mM to about 100 mM.10. The nucleic acid preservative according to claim 1 , wherein the chelating agent is EDTA and wherein the ...

Bicyclic nucleosides and oligomeric compounds prepared therefrom

The present invention provides novel 3′,5′-linked bicyclic nucleosides and oligomeric compounds prepared therefrom. The bicyclic nucleosides provided herein are useful for enhancing one or more properties of the oligomeric compounds they are incorporated into such as nuclease resistance.

Mechanically-Activated Cation Channels

Methods of screening for agents that modulate the activity of a mechanically-activated cation channel are provided. Also provided are compositions and methods for ameliorating pain by antagonizing or inhibiting mechanically-activated cation channels. 145-. (canceled)46. A method of ameliorating pain in a subject , the method comprising administering to the subject a) an antibody that antagonizes the activity of a mechanically activated cation channel polypeptide of SEQ ID NO:2 , SEQ ID NO:4 , SEQ ID NO:18 , or SEQ ID NO:20 , or b) an antisense oligonucleotide or small interfering RNA (siRNA) complementary to at least 15 contiguous nucleotides of SEQ ID NOs 1 , 3 , 17 , or 19 wherein the antisense oligonucleotide or siRNA inhibits production of the mechanically activated cation channel polypeptide.471. The method of claim , wherein the polypeptide comprises SEQ ID NO:4 or SEQ ID NO:20.481. The method of claim wherein the polypeptide is expressed in bladder , colon , kidney , lung , or skin.491. The method of claim , wherein the polypeptide is expressed in a dorsal root ganglion neuron.501. The method of claim , wherein the subject is a mammal.511. The method of claim , wherein the subject is a human.521. The method of claim , wherein the pain is selected from the group consisting of acute mechanical pain , chronic mechanical pain , mechanical hyperalgesia , mechanical allodynia , arthritis , inflammation , dental pain , cancer pain , and labor pain.531. The method of claim , wherein the antibody is a monoclonal antibody , a humanized antibody or a chimeric antibody.541. The method of claim , wherein the antisense oligonucleotide or siRNA comprises any one of SEQ ID NOs 5-16.55. An isolated antisense oligonucleotide or small interfering RNA (siRNA) complementary to at least 15 contiguous nucleotides of SEQ ID NOs:1 , 3 , 17 , or 19 and encodes a mechanically-activated cation channel polypeptide , wherein the antisense oligonucleotide or siRNA inhibits production of ...

Method and Kit for Sequential Isolation of Nucleotide Species From a Sample

The invention provides a process and kit for serial isolation of DNA and RNA from the same sample. First, a siliceous solid support with preferential affinity for DNA over RNA is used to capture DNA in a lysate of a sample. Next, a siliceous solid support with similar affinity for RNA and DNA is used to capture RNA from the same lysate. The respective solid supports are recovered independent of each other, washed, and their bound nucleotide species are eluted. The invention further provides DNA and RNA prepared using the process in a minimal number of steps employing a minimal number of reagents. As the invention yields DNA and RNA of high quality and is amenable to automation, the invention may be used widely in the healthcare and pharmaceutical industries. 1. A method of serial recovery of two different nucleic acid species from a biological sample , comprising:in a first binding step, selectively binding a first nucleic acid species to a first solid phase by contacting the biological sample with the first solid phase that selectively binds the first nucleic acid species;separating the first solid phase with the bound first nucleic acid species from an unbound portion of the biological sample;in a second binding step, binding a second nucleic acid species to a second solid phase, different from the first solid phase, by contacting an unbound portion of the biological sample with the second solid phase that binds the first and second nucleic acid species;enzymatic digestion of the first nucleic acid species bound to the second solid phase; andisolating the second nucleic acid species from the second solid phase.2. The method according to claim 1 , further comprising:in the first binding step, homogenization and enzymatic digestion of the biological sample in a lysis buffer, prior to the selectively binding, thereby generating a lysate comprising the first nucleic acid species.3. The method according to claim 1 , wherein the biological sample is lysed in a lysis ...

MODULATION OF EIF4E EXPRESSION

Oligomeric compounds, compositions and methods are provided for modulating the expression of eIF4E. The antisense compounds may be single- or double-stranded and are targeted to nucleic acid encoding eIF4E. Methods of using these compounds for modulation of eIF4E expression and for diagnosis and treatment of diseases and conditions associated with expression of eIF4E are provided. 1. An antisense oligonucleotide consisting of 12 to 30 linked nucleosides and having a nucleobase sequence at least 95% complementary within the 3′ untranslated region (3′UTR) of a nucleic acid encoding human eIF4E , wherein the antisense oligonucleotide comprises at least one chemically modified sugar moiety , internucleoside linkage , or nucleobase.2. The antisense oligonucleotide of claim 1 , wherein the chemically modified sugar moiety is a 2′-O-(2-methoxyethyl) sugar moiety.3. The antisense oligonucleotide of claim 1 , wherein the chemically modified sugar moiety is a bicyclic sugar moiety.4. The antisense oligonucleotide of claim 3 , wherein the bicyclic sugar moiety comprises a 4′-(CH2)-O-2′ or 4′-(CH2)2-O-2′ group.5. The antisense oligonucleotide of claim 1 , wherein the modified internucleoside linkage is a phosphorothioate internucleoside linkage.6. The antisense oligonucleotide of claim 1 , wherein the modified nucleobase is a 5-methylcytosine.7. The antisense oligonucleotide of claim 1 , wherein the antisense oligonucleotide is chimeric.8. A pharmaceutical composition comprising the antisense oligonucleotide of claim 1 , and a pharmaceutically or physiologically acceptable carrier claim 1 , diluent claim 1 , or excipient.9. The pharmaceutical composition of claim 8 , which is in a form suitable for parenteral administration.10. The pharmaceutical composition of claim 9 , wherein said parenteral administration is via intravenous injection or infusion.11. The antisense oligonucleotide of claim 1 , wherein the antisense oligonucleotide has a nucleobase sequence at least 95% ...

OLIGONUCLEOTIDE SPECIFIC UPTAKE OF NANOCONJUGATES

The present invention concerns nanoparticles functionalized with an oligonucleotide and a domain for a variety of uses, including but not limited to gene regulation. More specifically, the disclosure provides a nanoparticle that is taken up by a cell at an efficiency different than a nanoparticle functionalized with the same oligonucleotide but does not contain a domain. 1. A nanoparticle functionalized with an oligonucleotide and a domain , the nanoparticle having the property of being taken up by a cell at an efficiency different than a nanoparticle functionalized with the same oligonucleotide but lacking the domain.2. The nanoparticle of wherein the domain is located 5′ to the oligonucleotide.3. The nanoparticle of wherein the domain is located 3′ to the oligonucleotide.4. The nanoparticle of wherein the domain is located at an internal region within the oligonucleotide.5. The nanoparticle of wherein the domain is colinear with the oligonucleotide.6. The nanoparticle of functionalized with a second oligonucleotide and the domain is associated with the second oligonucleotide.7. The nanoparticle of wherein the domain comprises a polythymidine (polyT) sequence comprising more than one thymidine residue.8. The nanoparticle of wherein the domain comprises a polythymidine (polyT) sequence comprising two thymidine residues.9. The nanoparticle of wherein the domain comprises a polythymidine (polyT) sequence comprising two claim 1 , three claim 1 , four claim 1 , five claim 1 , six claim 1 , seven claim 1 , eight claim 1 , nine claim 1 , ten claim 1 , eleven claim 1 , twelve claim 1 , thirteen claim 1 , fourteen claim 1 , fifteen claim 1 , sixteen claim 1 , seventeen claim 1 , eighteen claim 1 , nineteen claim 1 , or twenty thymidine residues.10. The nanoparticle of wherein the domain comprises a phosphate polymer (C3 residue).11. The nanoparticle of wherein the domain comprises two or more phosphate polymers (C3 residues).12. A method of modulating cellular uptake ...

MICROORGANISM NUCLEIC ACID PURIFICATION FROM HOST SAMPLES

The present disclosure provides systems, devices, and methods for purifying microorganism nucleic acid from a host sample, such as a whole blood sample from a human. In certain embodiments, devices and systems with multiple filters are employed and provide for the selective removal of blood cells and host nucleic acids from a sample in order to enrich for microorganism nucleic acid. 1. A system or device for purifying microorganism nucleic acid from a sample comprising:i) a first size-exclusion filter that: excludes passage therethrough of lymphocytes, monocytes, neutrophils, eosinophils, and basophils, and allows passage therethrough of erythrocytes, lymphocytes, and platelets; a) a second size-exclusion filter that: excludes passage therethrough of erythrocytes, lymphocytes, and platelets, and allows passage therethrough of microorganisms smaller than platelets; or', I) a second size-exclusion filter that excludes passage therethrough of erythrocytes and lymphocytes, but allows passage therethrough of platelets and microorganisms smaller than platelets, and', 'II) a third size-exclusion filter that excludes passage therethrough of platelets, but allows passage therethrough of microorganisms smaller than platelets; and, 'b) both of the following], 'ii) eitheriii) a shearing filter that mechanically shears said microorganisms such that microorganism nucleic acid is released.2. The system or device of claim 1 , further comprising iv) an affinity capture filter comprising affinity binding molecules that bind human or animal nucleic acid.3. The system or device of claim 2 , wherein said affinity binding molecules bind human repetitive nucleic acid sequences.4. The system or device of claim 1 , wherein said first size-exclusion filter has a pore size of about 10 μm.5. The system or device of claim 1 , wherein the second size-exclusion filter of (ii) (a) has a pore size of about 2 μm or wherein the second size-exclusion filter of (ii) (b) has a pore size of about 5 μm ...

ANTISENSE NUCLEIC ACIDS

The present invention provides a pharmaceutical composition which causes skipping of the 53rd exon in the human dystrophin gene with a high efficiency. 1. An antisense oligomer which causes skipping of the 53rd exon in the human dystrophin gene , consisting of a nucleotide sequence complementary to any one of the sequences consisting of the 32nd to the 56th or the 36th to the 56th nucleotides from the 5′ end of the 53rd exon in the human dystrophin gene.2. The antisense oligomer according to claim 1 , which is an oligonucleotide.3. The antisense oligomer according to claim 2 , wherein the sugar moiety and/or the phosphate-binding region of at least one nucleotide constituting the oligonucleotide is modified.4. The antisense oligomer according to claim 3 , wherein the sugar moiety of at least one nucleotide constituting the oligonucleotide is a ribose in which the 2′-OH group is replaced by any one selected from the group consisting of OR claim 3 , R claim 3 , R′OR claim 3 , SH claim 3 , SR claim 3 , NH claim 3 , NHR claim 3 , NR claim 3 , N claim 3 , CN claim 3 , F claim 3 , Cl claim 3 , Br and I (wherein R is an alkyl or an aryl and R′ is an alkylene).5. The antisense oligomer according to claim 3 , wherein the phosphate-binding region of at least one nucleotide constituting the oligonucleotide is any one selected from the group consisting of a phosphorothioate bond claim 3 , a phosphorodithioate bond claim 3 , an alkylphosphonate bond claim 3 , a phosphoramidate bond and a boranophosphate bond.6. The antisense oligomer according to claim 1 , which is a morpholino oligomer.7. The antisense oligomer according to claim 6 , which is a phosphorodiamidate morpholino oligomer.911-. (canceled)12. The antisense oligomer according to claim 1 , consisting of the nucleotide sequence shown by SEQ ID NO: 11 or 35.13. A pharmaceutical composition for the treatment of muscular dystrophy claim 1 , comprising as an active ingredient the antisense oligomer according to claim 1 , or ...

OLIGONUCLEOTIDE AND USE THEREOF

Provided is an oligonucleotide containing an azobenzene derivative, represented by Formula (1) or (2) below: 2. The oligonucleotide according to claim 1 , wherein Rto Rand Rto Rare hydrogen atoms.3. The oligonucleotide according to claim 1 , wherein Rand Rare methyl groups.5. The oligonucleotide according to claim 4 , wherein Rand Rbind with each other to represent a cyclohexyl group or adamantyl group together with a carbon atom for linking to a sulfur atom.6. The oligonucleotide according to claim 4 , wherein{'sup': 31', '32, 'sub': '1-4', 'Rand Reach independently represent a Calkyl group, and'}{'sup': 33', '40, 'Rto Rare hydrogen atoms.'}7. The oligonucleotide according to claim 6 , wherein Rand Rare methyl groups.8. A photo-switching agent claim 1 , by which the formation and dissociation of a double strand can be controlled by visible light irradiation claim 1 , and which is provided with the oligonucleotide according to .10. The photo-switching agent according to claim 9 , wherein Rand Rrepresent methyl groups or hydrogen atoms claim 9 , and R claim 9 , R claim 9 , and Rto Rare hydrogen atoms.11. The photo-switching agent according to claim 9 , wherein each of the pair of oligonucleotides has two or more of the azobenzene derivative adjacent to one another on either side of two or more nucleotides. The present invention relates to an oligonucleotide, and to a photo-switching agent using the oligonucleotide. The priority claim for the present application is based on Japanese Patent Application No. 2010-194942, submitted on Aug. 31, 2010, and the entire contents of the Description of that Japanese Patent Application are incorporated by reference in this Description.Techniques have been developed for controlling hybridization between oligonucleotides with complementary structures by an external stimulus. If hybridization control could be achieved, it could contribute to more precise gene detection, identification and assay, and to the development of molecular ...

BUILDING BLOCKS AND METHODS FOR THE SYNTHESIS OF 5-HYDROXYMETHYLCYTOSINE-CONTAINING NUCLEIC ACIDS

The present invention relates to building blocks and methods for the efficient synthesis of 5-hydroxymethylcytosine-containing nucleic acids such as DNA or RNA. 2. The compound of claim 1 , wherein Z is a 5- or 6-membered cyclic radical claim 1 , particularly a ribose claim 1 , ribose analogue or deoxyribose radical claim 1 , wherein the 3′-OH group of the ribose claim 1 , ribose analogue or deoxyribose radical may be substituted by a phosphor-containing group claim 1 , e.g. a phosphate claim 1 , phosphoester or phosphoramidite group and wherein the 5′-OH group of the ribose claim 1 , ribose analogue or deoxyribose radical may be substituted by a protection group.4. The compound of claim 3 , wherein Ris an aliphatic linear or cyclic group comprising up to 6 C-atoms and optionally up to 2 heteroatoms such as N or O claim 3 , e.g. a C(halo) alkyl group claim 3 , or a C(hetero) alkyl group claim 3 , or a Caryl or heteroaryl group optionally substituted by OH claim 3 , halo claim 3 , CN claim 3 , (O)C(halo) alkyl or N(R) claim 3 , wherein Ris as defined for the compound of formula (II).5. A method of introducing a formyl substituent at position 5 of a cytosine cytidine claim 3 , or deoxycytidine compound comprising reacting a 5-halo substituted starting compound with CO under catalysis of Pd.6. A method for the synthesis of a nucleic acid claim 1 , comprising incorporating a compound of into said nucleic acid.7. A method of claim 6 , wherein the nucleic acid synthesis is carried out by a phosphoramidite procedure.8. A nucleic acid molecule having incorporated at least one compound of . The present invention relates to building blocks and methods for the efficient synthesis of 5-hydroxymethylcytosine-containing nucleic acids such as DNA or RNA.The genetic material is constructed from the four canonical bases dA, dC, dG, and dT. The dC base is furthermore subject to epigenetic modification. In eucaryotes the dC base is often methylated at position C5 to give the base 5- ...

LINKERS AND CO-COUPLING AGENTS FOR OPTIMIZATION OF OLIGONUCLEOTIDE SYNTHESIS AND PURIFICATION ON SOLID SUPPORTS

A method of modulation of synthesis capacity on and cleavage properties of synthetic oligomers from solid support is described. The method utilizes linker molecules attached to a solid surface and co-coupling agents that have similar reactivities to the coupling compounds with the surface functional groups. The preferred linker molecules provide an increased density of polymers and more resistance to cleavage from the support surface. The method is particularly useful for synthesis of oligonucleotides, oligonucleotides microarrays, peptides, and peptide microarrays. The stable linkers are also coupled to anchor molecules for synthesis of DNA oligonucleotides using on support purification, eliminating time-consuming chromatography and metal cation presence. Oligonucleotides thus obtained can be directly used for mass analysis, DNA amplification and ligation, hybridization, and many other applications. 1. A compound of the structure:{'br': None, 'sub': s', 'p, 'R-L-R'}{'sub': s', 'p, 'wherein Ris a substrate attaching group, Ris a polymer attaching group, and L is a linker.'}2. The compound of claim 1 , wherein said substrate attaching group is selected from the group consisting of chlorosilyl and alkyloxysilyl functional groups.3. The compound of claim 1 , wherein said polymer attaching group is selected from the group consisting of amine claim 1 , hydroxyl claim 1 , thiol claim 1 , carboxylic acid claim 1 , ester claim 1 , amide claim 1 , epoxide claim 1 , isocyanate claim 1 , and isothiocyanate.4. A method for controlling the number of oligonucleotides synthesized at a predetermined site on a substrate comprising: i) a substrate;', 'ii) a plurality of non-cleavable linkers;', {'sub': '2', 'iii) a plurality of anchor moieties, wherein said anchor moieties comprise the structure —C(X)—C(Y), wherein X comprises —OPOO—, Y is a nucleophile and wherein said structure is part of a ring moiety;'}, 'iv) nucleotide monomers; and', 'v) co-coupling agents;, 'a) providingb) ...

DNA SEQUENCING BY SYNTHESIS USING MODIFIED NUCLEOTIDES AND NANOPORE DETECTION

This disclosure is related to a method of sequencing a single-stranded DNA using deoxynucleotide polyphosphate analogues and translocation of tags from incorporated deoxynucleotide polyphosphate analogues through a nanopore. 5. The method of claim 1 , wherein the tag is ethylene glycol claim 1 , an amino acid claim 1 , a carbohydrate claim 1 , a dye claim 1 , a mononucleotide claim 1 , a dinucleotide claim 1 , a trinucleotide claim 1 , a tetranucleotide claim 1 , a pentanucleotide or a hexanucleotide claim 1 , a fluorescent dyes claim 1 , a chemiluminiscent compound claim 1 , an amino acid claim 1 , a peptide claim 1 , a carbohydrate claim 1 , a nucleotide monophopshate claim 1 , a nucleotide diphosphate claim 1 , an aliphatic acid or an aromatic acid or an alcohol or a thiol with unsubstituted or substituted with one or more halogens claim 1 , a cyano group claim 1 , a nitro group claim 1 , an alkyl group claim 1 , an alkenyl group claim 1 , an alkynyl group claim 1 , an azido group.6. The method of claim 1 , wherein the base is selected from the group consisting of adenine claim 1 , guanine claim 1 , cytosine claim 1 , thymine claim 1 , 7-deazaguanine claim 1 , 7-deazaadenine or 5-methylcytosine.7. The method of claim 1 , further comprising a washing step after each iteration of step (b) to remove unincoporated dNPP analogues from contact with the single-stranded DNA.8. (canceled)9. The method of claim 1 , wherein the single-stranded DNA claim 1 , electrolyte solution and nanopore in the membrane are located within a single container.1011-. (canceled)13. The method of claim 1 , wherein the tag is a mononucleotide claim 1 , a dinucleotide claim 1 , a trinucleotide claim 1 , a tetranucleotide claim 1 , a pentanucleotide or a hexanucleotide and wherein the base of the mononucleotide claim 1 , a dinucleotide claim 1 , a trinucleotide claim 1 , a tetranucleotide claim 1 , a pentanucleotide or a hexanucleotide is the same type of base as the base of the dNPP analogue. ...

OLIGONUCLEOTIDE WITH PROTECTED BASE

The present invention provides a protected nucleotide for elongation, which can be purified efficiently and in a high yield by a liquid-liquid extraction operation, and can achieve an oligonucleotide production method by a phosphoramidite method. 2. The oligonucleotide comprising a protected base of according to claim 1 , wherein q is 0.4. The oligonucleotide comprising a protected base according to claim 3 , wherein R claim 3 , R claim 3 , Rin the number of l claim 3 , Rand Rare each independently a branched chain alkyl group or branched chain alkenyl group selected from the group consisting of a 2 claim 3 ,6 claim 3 ,10 claim 3 ,14-tetramethylpentadecyl group claim 3 , a 2 claim 3 ,6 claim 3 ,10-trimethylundecyl group claim 3 , a 2 claim 3 ,2 claim 3 ,4 claim 3 ,8 claim 3 ,10 claim 3 ,10-hexamethyl-5-undecyl group claim 3 , a 2 claim 3 ,6 claim 3 ,10-trimethylundeca-1 claim 3 ,5 claim 3 ,9-trienyl group claim 3 , a 2 claim 3 ,6-dimethylheptyl group claim 3 , a 2 claim 3 ,6-dimethylhept-5-enyl group claim 3 , a 2 claim 3 ,6-dimethylhepta-1 claim 3 ,5-dienyl group claim 3 , a 9-nonadecyl group claim 3 , a 12-methyltridecyl group claim 3 , an 11-methyltridecyl group claim 3 , an 11-methyldodecyl group claim 3 , a 10-methylundecyl group claim 3 , an 8-heptadecyl group claim 3 , a 7-pentadecyl group claim 3 , a 7-methyloctyl group claim 3 , a 3-methyloctyl group claim 3 , a 3 claim 3 ,7-dimethyloctyl group claim 3 , a 3-methylheptyl group claim 3 , a 3-ethylheptyl group claim 3 , a 5-undecyl group claim 3 , a 2-heptyl group claim 3 , a 2-methyl-2-hexyl group claim 3 , a 2-hexyl group claim 3 , a 3-heptyl group claim 3 , a 4-heptyl group claim 3 , a 4-methyl-pentyl group claim 3 , a 3-methyl-pentyl group claim 3 , and a 2 claim 3 ,4 claim 3 ,4-trimethylpentyl group; or a straight chain alkyl group selected from the group consisting of a tetradecyl group claim 3 , a tridecyl group claim 3 , a dodecyl group claim 3 , an undecyl group claim 3 , a decyl group claim 3 , a ...

Synthesis of oligonucleotides

A method for preparing an oligonucleotide comprising the steps of synthesizing a phosphoramidite by reacting a hydroxyl-containing compound of formula (A) with a phosphitylating agent in the presence of an activator compound of formula (I), to prepare a phosphitylated compound, then coupling the phosphitylated compound without isolation with a second compound having the formula (A), wherein R, R, R, B are independently selected, but have the same definition as above in the presence of an activator II selected from the group of imidazole, imidazolium salts, and mixtures thereof, which are improved activators over activators disclosed in related art. 4. The method of claim 1 , wherein the phosphitylating agent is 2-cyanoethyl-N claim 1 ,N claim 1 ,N′ claim 1 ,N′-tetraisopropylphosphorodiamidite.5. The method of claim 1 , wherein the deprotonated acid is selected from the group consisting of trifluoroacetic acid claim 1 , dichloroacetic acid claim 1 , methanesulfonic acid claim 1 , trifluoromethanesulfonic acid claim 1 , and o-chlorophenolic acid.6. The method of claim 1 , wherein the reacting is in the presence of acetone.7. The method of claim 1 , wherein the concentration of phosphitylating agent in step b) is from 1.0 to 1.2 mol/mol of hydroxyl groups in the hydroxyl containing compound.8. The method of claim 1 , wherein the concentration of phosphitylating agent in step b) is from 3 to 5 mol/mol of hydroxyl groups in the hydroxyl containing compound.9. The method of claim 1 , further comprising adding a polymeric alcohol after step b).10. The method of claim 9 , wherein the polymeric alcohol is polyvinyl alcohol.11. The method of claim 1 , wherein the deprotonated acid is selected from the group consisting of trifluoroacetic acid claim 1 , dichloroacetic acid claim 1 , methanesulfonic acid claim 1 , trifluormethanesulfonic acid (triflate) claim 1 , o-chlorophenolate claim 1 , and mixtures thereof.12. The method of claim 9 , wherein the reacting is in the presence ...

METHODS FOR TREATING MULTIPLE SCLEROSIS USING ANTISENSE OLIGONUCLEOTIDES

A method for treating a patient suffering from multiple sclerosis, particularly a relapsing form of multiple sclerosis, comprising periodically administering a pharmaceutical composition comprising a therapeutically effective amount of OLIGONUCLEOTIDE 1 to the patient, thereby treating the patient. 1. A method for decreasing or inhibiting the accumulation of new active brain lesions , or decreasing or inhibiting an increase in the volume of gadolinium-enhancing brain lesions in a human subject afflicted with multiple sclerosis , comprising periodically administering to the human subject a pharmaceutical composition comprising a pharmaceutically acceptable carrier and a therapeutically effective amount of an oligonucleotide having the structure:{'br': None, 'sup': Me', 'Me', 'Me', 'Me', 'Me', 'Me', 'Me', 'Me', 'Me', 'Me, '5′-CUG AGT CTG TTT UCC AUU CU-3′'} a) each of the 19 internucleotide linkages of the oligonucleotide is an O,O-linked phosphorothioate diester;', 'b) the nucleotides at the positions 1 to 3 from the 5′ end are 2′-O-(2-methoxyethyl) modified ribonucleosides;', 'c) the nucleotides at the positions 4 to 12 from the 5′ end are 2′-deoxyribonucleosides;', 'd) the nucleotides at the positions 13 to 20 from the 5′ end are 2′-O-(2-methoxyethyl) modified ribonucleosides; and', {'sup': 'Me', 'e) all cytosines are 5-methylcytosines (C),'}], 'wherein'}or a pharmaceutically acceptable salt of the oligonucleotide, so as to thereby decrease or inhibit the accumulation of new active brain lesions, or decrease or inhibit an increase in the volume of gadolinium-enhancing brain lesions in a human subject afflicted with multiple sclerosis.2. The method of claim 1 , wherein the method inhibits progression of disability in the human subject or reduces relapse rate in the human subject.3. The method of claim 95 , wherein the progression of disability is reduced by 15%-70% as measured by EDSS score relative to the progression of disability in a human subject afflicted with ...

Methods and kits for breaking emulsions

The disclosure relates generally to methods, systems, compositions and kits for breaking a water-in-oil emulsion including one or more biomolecules dispersed in an aqueous phase of the water-in-oil emulsion. In some embodiments, the disclosure relates to obtaining a first emulsion including a continuous hydrophobic fraction and a discontinuous aqueous fraction, the aqueous fraction having one or more biomolecules dispersed therein, breaking the first emulsion by contacting the first emulsion with a breaking solution including a second emulsion, where the second emulsion includes a discontinuous phase of organic extraction solvent dispersed in a continuous aqueous phase, and centrifuging to separate the phases of the resulting mixture. In some embodiments, the disclosure relates generally to methods, kits and systems for extracting biomolecules from a water-in-oil emulsion, including breaking a water-in-oil emulsion comprising a plurality of aqueous droplets in a continuous hydrophobic fraction using a breaking solution to produce a resulting reaction mixture containing one or more biomolecules and manipulating the resulting reaction mixture to form at least two phases, where one of the phases includes an aqueous phase containing the one or more biomolecules.

ALIGNMENT OF NANOMATERIALS AND MICROMATERIALS

The present invention provides a method for preparing a nanoassembly that includes the step of reacting the assembly template with at least one nanomaterial to form the nanoassembly using a bifunctional linker. 146.-. (canceled)47. A method of generating an assembly with a desired linear , two-dimensional or three-dimensional structure , comprising:(a) selecting one or more internal positions of a nucleic acid polymer template for attachment of a particle;(b) predicting a linear, two-dimensional and three-dimensional structure of the nucleic acid polymer template when the particle is attached to the one or more selected internal positions of the nucleic acid polymer; [ a single-stranded molecule or a double-stranded molecule comprising DNA, RNA, PNA, or mixed co-polymers thereof, and', 'one or more modified phosphodiester linkages at the selected one or more internal positions within the single stranded molecule or within one or both strands of the double-stranded molecule, wherein the one or more modified phosphodiester linkages each comprise a reactive substituent,, 'wherein the nucleic acid polymer template comprises, 'and wherein the particle comprises at least one linking reagent comprising a first reactive group and a second reactive group separated by a linker segment, wherein the second reactive group is attached the particle,', 'under conditions that permit the first reactive group to attach to the reactive substituent, and, '(c) reacting the nucleic acid polymer template with the particle,'}(d) coupling the particle to the one or more selected internal positions of the nucleic acid polymer template through the linking reagent, thereby forming the assembly with the desired linear, two-dimensional or three-dimensional structure.48. The method of claim 47 , further comprising synthesizing the nucleic acid polymer template claim 47 , wherein synthesizing comprises introducing the reactive substituents at the selected one or more internal positions.49. The ...

OLIGONUCLEOTIDE REPLACEMENT FOR DI-TAGGED AND DIRECTIONAL LIBRARIES

Transposomes and oligonucleotide replacement methods to make DNA libraries that have distinct 5′ and 3′ tags, and to make directional libraries that are enriched for a desired strand. 1. A method for adding a tag to the double-stranded product of a tagmentation reaction , comprising the steps of:(a) providing a double-stranded target nucleic acid and a transposome having a transposase with two transposon end sequences: a “transferred strand” and a “non-transferred strand”;(b) allowing the transposome to fragment the target nucleic acid, whereby the transferred strand is covalently transferred to a first strand of the fragment, and the non-transferred strand remains hybridized to the transferred strand;(c) removing the non-transferred strand from the transferred strand;(d) providing a replacement oligo that comprises a tag sequence, to hybridize to transferred strand; and(e) ligating the replacement oligo to the second strand of the fragment;thereby generating a tagmentation product having a transferred strand and a replacement oligo.2. The method of claim 1 , wherein one strand of the target nucleic acid is chemically modified.3. The method of claim 2 , wherein the chemical modification is conversion of cytosines to uracils.4. The method of claim 1 , wherein the transposome comprises a Tn5 transposase.5. The method of claim 1 , wherein the transposome comprises a Mu transposase.7. The method of claim 6 , wherein the transferred strand further comprises a tag sequence.9. The method of claim 1 , wherein the non-transferred strand is selected from the group consisting of SEQ ID NO:9 claim 1 , SEQ ID NO:10 claim 1 , SEQ ID NO:11 claim 1 , SEQ ID NO:12 claim 1 , SEQ ID NO:13 claim 1 , SEQ ID NO:14 claim 1 , SEQ ID NO:15 claim 1 , SEQ ID NO:16 claim 1 , SEQ ID NO:17 claim 1 , and SEQ ID NO:18.10. The method of claim 1 , wherein step (e) further comprises an extension step.11. The method of claim 2 , further comprising the step of(f) enriching for a desired strand.12. The ...

HYBRID SOLID SUPPORTS USEFUL FOR OLIGONUCLEOTIDE PRODUCTION

A method for preparing a crosslinked polymer coated controlled porosity glass (CPG) particle is provided. The method involves mixing CPG particles in a solution comprising polyvinylbenzylchloride and a first solvent at a temperature below 10° C. A second solvent is added and a crosslinking agent is added to the mixture. The first solvent is removed rapidly within 1½ hours of addition of the crosslinking agent. The crosslinking reaction is permitted to proceed and the mixture is then cooled and treated to remove any remaining solvent. The resulting coated CPG particles are washed and dried. Also provided a polymer coated CPG particles using for loading ligand thereon. 1. A conformal polymer-coated controlled porosity glass (CPG) particle for solid phase chemical synthesis comprising:(a) a controlled porosity glass core having pores with a mean average diameter of 500 Angstroms to 4000 Angstroms (50 nm to 4000 nm); and(b) a conformal cross-linked polymeric coating over said core, wherein said polymer comprises a cross-linking agent and monomeric subunits or a polymerizable monomer selected from the group consisting of a vinylbenzylchloride, an acrylic, a styrene, polymers thereof, and mixtures thereof,wherein the conformal polymeric coating layer is on the surface of the CPG particle and within its pores without changing the shape of the coated CPG particle.2. The conformal polymer-coated CPG particle according to claim 1 , wherein the pores within the conformal coated CPG particle retain an average pore size of at least 90% of the average pore size of the uncoated CPG particles (a) claim 1 , based on dry pore size.3. The conformal polymer-coated CPG particle according to claim 1 , wherein the polymeric coating adheres to the CPG core is the absence of a coupling agent to link the coating to the CPG particle.4. The conformal polymer-coated CPG particle according to claim 1 , wherein the polymeric coating is crosslinked in an amount of about 5% to about 30% crosslinked ...

COMPOUNDS AND METHODS FOR MODULATING ANGIOTENSINOGEN EXPRESSION

Disclosed herein are compositions and compounds comprising modified oligonucleotides for modulating AGT and modulating a RAAS pathway related disease, disorder and/or condition in an individual in need thereof. A RAAS pathway related disease, disorder and/or condition in an individual such as hypertension can be treated, ameliorated, delayed or prevented with the administration of antisense compounds targeted to AGT. 1. An oligomeric compound comprising a modified oligonucleotide consisting of 12 to 30 linked nucleosides having a nucleobase sequence comprising a portion of at least 8 contiguous nucleobases complementary to an equal length portion of nucleobases 525 to 560 of SEQ ID NO: 1 , wherein the nucleobase sequence of the modified oligonucleotide is at least 80% complementary to SEQ ID NO: 1.2. An oligomeric compound comprising a modified oligonucleotide consisting of 12 to 30 linked nucleosides having a nucleobase sequence comprising a portion of at least 8 contiguous nucleobases complementary to an equal length portion of nucleobases 643-691 of SEQ ID NO: 1 , wherein the nucleobase sequence of the modified oligonucleotide is at least 80% complementary to SEQ ID NO: 1.3. The oligomeric compound of claim 1 , wherein the modified oligonucleotide consists of 15 to 30 linked nucleosides.46-. (canceled)7. An oligomeric compound comprising a modified oligonucleotide comprising 12 to 30 linked nucleosides and having a nucleobase sequence comprising at least 19 contiguous nucleobases of the nucleobase sequence of SEQ ID NOs: 46 claim 1 , 53-54 claim 1 , 61 claim 1 , 68 claim 1 , 76 claim 1 , 83 claim 1 , 85 claim 1 , 93 claim 1 , 96-97 claim 1 , 109 claim 1 , 134-135 claim 1 , 137-39 claim 1 , 142 claim 1 , 163-172 claim 1 , 180-184 claim 1 , 186 claim 1 , 189 claim 1 , 234 claim 1 , 236 claim 1 , 238-239 claim 1 , 267 claim 1 , 313 claim 1 , 411 claim 1 , 452 claim 1 , 463-470 claim 1 , 475-478 claim 1 , 480 claim 1 , 500-503 claim 1 , 512 claim 1 , 517-518 claim 1 ...

MODULATION OF FACTOR 11 EXPRESSION

Disclosed herein are antisense compounds and methods for decreasing Factor 11 and treating or preventing thromboembolic complications in an individual in need thereof. Examples of disease conditions that can be ameliorated with the administration of antisense compounds targeted to Factor 11 include thrombosis, embolism, and thromboembolism, such as, deep vein thrombosis, pulmonary embolism, myocardial infarction, and stroke. Antisense compounds targeting Factor 11 can also be used as a prophylactic treatment to prevent individuals at risk for thrombosis and embolism. 154.-. (canceled)55. A compound comprising a modified oligonucleotide , wherein the modified oligonucleotide consists of 12 to 30 linked nucleosides , and wherein at least 12 linked nucleosides of the modified oligonucleotide has a nucleobase sequence of SEQ ID NO: 141.56. The compound of claim 55 , wherein the modified oligonucleotide has a nucleobase sequence that is at least 85% complementary to any of the nucleobase sequences of SEQ ID NO: 1 or SEQ ID NO: 2 when measured across the entire nucleobase sequence of the modified oligonucleotide.57. The compound of claim 55 , wherein the modified oligonucleotide consists of a single stranded modified oligonucleotide.58. The compound of claim 55 , wherein the modified oligonucleotide comprises at least one modified internucleoside linkage.59. The compound of claim 58 , wherein the at least one modified internucleoside linkage is a phosphorothioate internucleoside linkage.60. The compound of claim 55 , wherein the modified oligonucleotide comprises at least one modified sugar.61. The compound of claim 60 , wherein the at least one modified sugar comprises a 2′-O-methoxyethyl group.62. The compound of claim 60 , wherein the at least one modified sugar comprises a 2′-O—CHgroup.63. The compound of claim 60 , wherein the at least one modified sugar is a bicyclic sugar.64. The compound of claim 63 , wherein the bicyclic sugar comprises a 4′-(CH)—O-2′ bridge.65. ...

TRICYCLIC NUCLEOSIDES AND OLIGOMERIC COMPOUNDS PREPARED THEREFROM

The present invention provides fluorine substituted tricyclic nucleosides of formula (I) wherein: the substituents are as defined in the claims. The present invention further provides oligomeric compounds prepared therefrom. Incorporation of one or more of the tricyclic nucleosides into an oligomeric compound enhances one or more properties of the oligomeric compound. Such oligomeric compounds can also be included in double stranded compositions. 2. The tricyclic nucleoside of claim 1 , wherein Bx is uracil claim 1 , thymine claim 1 , cytosine claim 1 , 5-methylcytosine claim 1 , adenine or guanine.3. The tricyclic nucleoside of any one of the preceding claims claim 1 , wherein Tis hydroxyl or protected hydroxyl claim 1 , and wherein Tis a reactive phosphorus group selected from an H-phosphonate or a phosphoramidite.4. The tricyclic nucleoside of any one of the preceding claims claim 1 , wherein Tis 4 claim 1 ,4′-dimethoxytrityl and Tis diisopropylcyanoethoxy phosphoramidite or a controlled pore glass surface.5. The tricyclic nucleoside of any one of the preceding claims claim 1 , wherein one or two of q claim 1 , q claim 1 , qzand zis F claim 1 , and the other of q claim 1 , q claim 1 , qzand zare H.8. The oligomeric compound of claims 7 , wherein each Bx is claims 7 , independently claims 7 , uracil claims 7 , thymine claims 7 , cytosine claims 7 , 5-methylcytosine claims 7 , adenine or guanine.9. The oligomeric compound of any one of to claims 7 , wherein each internucleoside linking group is claims 7 , independently claims 7 , a phosphodiester internucleoside linking group or a phosphorothioate internucleoside linking group.10. The oligomeric compound of any one of to claims 7 , wherein essentially each internucleoside linking group is a phosphorothioate internucleoside linking group.11. The oligomeric compound of any one of to claims 7 , comprising a first region having at least two contiguous tricyclic nucleosides having Formula II.12. The oligomeric compound ...

Novel Compounds

A compound of Formula (I) 2. The method of wherein the cancer is melanoma.3. The method of wherein the cancer is head and neck cancer.4. The method of wherein the compound is administered by intratumoral injection.5. The method of wherein the compound is administered by peritumoral injection.6. The method of wherein the cancer is squamous cell carcinomas.7. The method of wherein the cancer is breast cancer.8. The method of wherein the compound is administered by intratumoral injection.9. The method of wherein the compound is administered by peritumoral injection.10. The method of wherein the cancer is hepatocellular cancer.11. The method of wherein the cancer is colon cancer.12. The method of wherein the cancer is esophageal cancer.13. The method of wherein the cancer is rectal cancer.14. The method of wherein the cancer is lung cancer.15. The method of wherein the compound is administered by intratumoral injection.16. The method of wherein the compound is administered by peritumoral injection.17. The method of wherein the cancer is renal cell cancer.18. The method of wherein claim 1 , the cancer is selected from: melanoma claim 1 , squamous cell carcinomas claim 1 , hepatocellular cancer claim 1 , colon cancer claim 1 , esophageal cancer claim 1 , rectal cancer claim 1 , and renal cell cancer claim 1 , and the compound is administered by intratumoral injection.19. The method of claim 1 , the wherein claim 1 , the cancer is selected from: melanoma claim 1 , squamous cell carcinomas claim 1 , hepatocellular cancer claim 1 , colon cancer claim 1 , esophageal cancer claim 1 , rectal cancer claim 1 , and renal cell cancer claim 1 , and the compound is administered by peritumoral injection.20. The method of wherein the cancer is selected from: brain (gliomas) claim 1 , glioblastomas claim 1 , astrocytomas claim 1 , glioblastoma multiforme claim 1 , Bannayan-Zonana syndrome claim 1 , Cowden disease claim 1 , Lhermitte-Duclos disease claim 1 , Wilm's tumor claim 1 , Ewing' ...

Flex plate with removable inserts and cover

Technologies are described for methods and systems effective for flex plates. The flex plates may comprise a base plate. The base plate may include walls that define an insert location opening in the base plate. The insert location opening in the base plate may be in communication with a securement area. The flex plates may comprise an insert. The insert may include a reservoir region and a crystallization region separated by a wall including channels. The reservoir region and the crystallization region may include a backing. The insert may further include securement tabs. The securement tabs may be configured to secure the insert to the base plate at the securement area.

A METHOD FOR GENERATING RANDOM OLIGONUCLEOTIDES AND DETERMINING THEIR SEQUENCE

Random oligonucleotides are generated with incomplete information about the sequence of the nucleic acid bases present in the newly generated molecules. The sequences of the oligonucleotides are subsequently determined and then these oligonucleotides can be processed for various potential uses. 1. A method of generating an oligonucleotide , the method comprising:a. generating at least one molecule comprising nucleotides by adding at least one nucleotide at random to the molecule, wherein the molecule generated is a random oligonucleotide;b. determining the sequence of the random oligonucleotide; andc. selecting random oligonucleotides using certain characteristics of the random oligonucleotides.2. The method of wherein the random oligonucleotides are generated using phosphoramidite chemistry.3. The method of claim 1 , wherein the random oligonucleotides are generated using an enzymatic process.4. The method of claim 1 , wherein the random oligonucleotides are generated within a microwell.5. The method of claim 1 , wherein the random oligonucleotides are generated on a microarray.6. The method of wherein the characteristic used to select the random oligonucleotide is a specific sequence of nucleotides.7. The method of wherein the characteristic to select the random oligonucleotide is a size of the random oligonucleotide.8. The method of wherein the random oligonucleotides are generated on oligonucleotides having a sequence that is at least partially known.9. The method of claim 3 , wherein an indicator molecule becomes reactive after a nucleotide is added to the molecule.10. The method of wherein adding a nucleic acid base to the molecules is partially directed.11. The method of claim 1 , wherein microfluids are used to control reaction conditions.12. The method of wherein directed energy is used to control the reaction conditions.13. The method of where the properties of the random oligonucleotide are measured using a nanopore.14. The method of wherein the selected ...

LIGHT-STIMULATED RELEASE OF CARGO FROM OLIGONUCLEOTIDES

The invention provides oligonucleotide conjugates including a photolabile crosslinker attached to a cargo moiety, e.g., a therapeutic or diagnostic agent. The invention further provides reagents useful in the preparation of such conjugates and methods of their use. 2. The conjugate of claim 1 , wherein L is present and amido claim 1 , C1-10 alkylene claim 1 , or C1-20 polyalkeneoxide.3. The conjugate of claim 1 , wherein L is present and C2-C20 polyethylene glycol.4. The conjugate of claim 1 , wherein Ais triazolyl claim 1 , disulfide claim 1 , cyclohexenyl claim 1 , amido claim 1 , thioamido claim 1 , acetal claim 1 , ketal claim 1 , or sulfonamido.5. The conjugate of claim 1 , wherein Y is C1-10 alkyl.6. The conjugate of claim 1 , wherein Y is methyl.7. The conjugate of claim 1 , wherein Ais the amine reactive group.8. The conjugate of claim 7 , wherein the amine reactive group is p-nitrophenoxyl claim 7 , N-hydroxysuccinimidyl claim 7 , halide claim 7 , pentafluorophenoxyl claim 7 , or imidazolyl.9. The conjugate of claim 1 , wherein Ais —NHX.10. The conjugate of claim 9 , wherein X is a therapeutic or diagnostic agent.13. The crosslinker of claim 12 , wherein L is present and C1-11 alkylene or C1-20 polyalkeneoxide.14. The crosslinker of claim 12 , wherein L is present and C2-C20 polyethylene glycol.15. The crosslinker of claim 12 , wherein Ais azido claim 12 , alkynyl claim 12 , alkenyl claim 12 , thiol claim 12 , halide claim 12 , boronic acid claim 12 , hydroxyl claim 12 , carboxyl claim 12 , formyl claim 12 , or ketone.16. The crosslinker of claim 12 , wherein Y is C1-11 alkyl.17. The crosslinker of claim 12 , wherein Y is methyl.18. The crosslinker of claim 12 , wherein Ais p-nitrophenoxyl claim 12 , N-hydroxysuccinimidyl claim 12 , halide claim 12 , pentafluorophenol claim 12 , or imidazolyl.20. An oligonucleotide conjugate comprising an oligonucleotide of 2-1000 nucleotides in length claim 12 , conjugated to a therapeutic or diagnostic agent by a ...

Mobile Solid Phase Reaction System and Method

A system and method are disclosed. A system for contacting a mobile solid phase with a flowing fluid phase includes: one or more reaction module, wherein the one or more reaction module comprises a conduit for the passage of a fluid phase and a solid phase, the conduit comprising a fluid input port and a fluid outlet port, and a first service module operably connected to a first side of a reaction module, the first service module for supplying and/or receiving the fluid phase to and/or from the reaction module, wherein the system is configured for passing a solid phase through the reaction module, via the conduit. 1. A system for contacting a mobile solid phase with a flowing fluid phase comprising: 'wherein the first reaction module comprises a conduit for the passage of a fluid phase and a solid phase, the conduit comprising a fluid input port and a fluid outlet port; and', 'a first reaction module;'}a first service module operably connected to a first side of the first reaction module, the first service module for supplying and/or receiving the fluid phase to and/or from the first reaction module;wherein the system is configured for passing a solid phase through the first reaction module, via the conduit.2. The system as claimed in further comprising a second reaction module claim 1 , provided in series claim 1 , such that the solid phase may pass through the consecutive reaction modules.3. The system as claimed in wherein the two reaction modules claim 2 , and the first service module claim 2 , are all configured to releasably connect to adjacent modules.4. The system as claimed in claim 1 , wherein the first side and a further side of the first reaction module are each a mating face; andwherein the first service module has a mating face that is connectable with a respective mating face of the first reaction module.5. The system as claimed in claim 1 , wherein the conduit of the first reaction module comprises a solid phase input port and a solid phase output ...

DIAMINOPHENOTHIAZINIUM DERIVATIVES FOR LABELLING BIOMOLECULES, METHOD AND SUBSTRATE FOR LABELLING OLIGONUCLEOTIDES, AND OLIGONUCLEOTIDES OBTAINED

The present invention relates to diaminophenothiazinium derivatives of formula (I); in which R, R, R, R, R, Rand X are as defined in Claim and also the methods for labelling oligonucleotides using such a derivative, labeling substrates and the oligonucleotides which can be obtained by means of such methods or from such labelling substrates. 2. The diaminophenothiazinium derivatives as claimed in claim 1 , characterized in that Aand Aare linear or branched alkylene chains in which from 2 to 6 consecutive carbon atoms separate the oxygen and nitrogen atoms.3. The diaminophenothiazinium derivatives as claimed in claim 1 , characterized in that at least one of the R claim 1 , R claim 1 , Rand Rgroups does not represent an -A-ORgroup as defined in and said R claim 1 , R claim 1 , Ror Rgroup(s) different than -A-ORand that -A-ORrepresent(s) an alkyl group having from 2 to 12 carbon atoms claim 1 , preferably from 4 to 12 carbon atoms.4. The diaminophenothiazinium derivatives as claimed in claim 1 , characterized in that R═R═H.5. The diaminophenothiazinium derivatives as claimed in claim 1 , characterized in that Rrepresents a —P{N[(C-C)alkyl]}(OCHCHC≡N) group claim 1 , such as the —P[N(Pr)](OCHCHC≡N) group.9. A method for labeling an oligonucleotide with a diaminophenothiazinium derivative as claimed in claim 1 , which comprises the growth of an oligonucleotide grafted onto a solid substrate claim 1 , and the replacement of one or more of the nucleotides of which it is formed with one or more of said diaminophenothiazinium derivatives claim 1 , before the oligonucleotide is detached from the solid substrate.10. The labeling method as claimed in claim 9 , characterized in that at least one replacement with a diaminophenothiazinium derivative is carried out before the end of the growth of the oligonucleotide.11. The labeling method as claimed in claim 9 , characterized in that at least one substitution with a diaminophenothiazinium derivative is carried out in the 3′ or 5′ ...

Modified L-Nucleic Acid

A modified L-nucleic acid, containing an L-nucleic acid part conjugated to a non-L-nucleic acid part is described. The conjugate has extended retention time in and demonstrates a delayed elimination from an organism. 1. (canceled)2. A modified L-nucleic acid , comprising a L-nucleic acid part and a non-L-nucleic acid part , wherein the L-nucleic acid part is conjugated with the non-L-nucleic acid part , and the conjugation of the L-nucleic acid part with the non-L-nucleic acid part leads to an increased retention time in an organism or a retarded excretion from an organism compared to a L-nucleic acid comprising only the L-nucleic acid part , and wherein said L-nucleic acid part is a spiegelmer.3. The modified L-nucleic acid of claim 2 , wherein the non-L-nucleic acid part has a molecular weight of more than about 300 Da.4. The modified L-nucleic acid of claim 2 , wherein the modified L-nucleic acid has a molecular weight of about 600 to 500 claim 2 ,000 Da.5. The modified L-nucleic acid of claim 2 , wherein the L-nucleic acid part has a molecular weight of 300 to 50 claim 2 ,000 Da.6. The modified L-nucleic acid of claim 2 , wherein the non-L-nucleic acid part is linked to the L-nucleic acid part via a functional group of the L-nucleic acid part claim 2 , wherein the functional group is selected from the group consisting of terminal and non-terminal phosphates claim 2 , terminal and non-terminal sugar portions claim 2 , natural and non-natural purine bases claim 2 , and natural and non-natural pyrimidine bases.7. The modified L-nucleic acid of claim 6 , wherein the linkage of the non-L-nucleic acid part with the L-nucleic acid part is via the 2′-OH— claim 6 , 3′-OH— claim 6 , 5′-OH-group or a derivative therefrom claim 6 , or one or more sugars of the L-nucleic acid part.8. The modified L-nucleic acid of claim 6 , wherein the linkage is via at least one of the positions 5 or 6 of a pyrimidine base.9. The modified L-nucleic acid of claim 6 , wherein the linkage is ...

Nucleic acid polyhedra from self-assembled vertex-containing fixed-angle nucleic acid structures

Provided herein are compositions comprising nucleic acid structures comprising three or more arms arranged at fixed angles from each other, composites thereof such as DNA cages, and methods for their synthesis and use.

Oligonucleotide Compositions and Methods of Making the Same

The present disclosure provides a solid phase method of making oligonucleotides via sequential coupling cycles including at least one coupling of a dinucleotide dimer subunit to a free 3′-terminal group of a growing chain. The oligonucleotides include at least two nucleoside subunits joined by a N3′→P5′ phosphoramidate linkage. The method may include the steps of (a) deprotecting the protected 3′ amino group of a terminal nucleoside attached to a solid phase support, said deprotecting forming a free 3′ amino group; (b) contacting the free 3′ amino group with a 3′-protected amino-dinucleotide-5′-phosphoramidite dimer in the presence of a nucleophilic catalyst to form an internucleoside N3′→P5′ phosphoramidite linkage; and (c) oxidizing (e.g., sulfurizing) the linkage. The compositions produced by the subject methods may include a reduced amount of one or more (N−x) oligonucleotide products. Also provided are pharmaceutical compositions including the subject oligonucleotide compositions. 141.-. (canceled)43. The method of claim 42 , wherein oxidizing the linkage comprises sulfurization to produce a thiophosphoramidate linkage.44. The method of claim 42 , wherein oxidizing the linkage produces an oxophosphoramidate linkage.45. The method of claim 42 , wherein the polynucleotide comprises a sequence of nucleoside subunits complementary to the RNA component of human telomerase claim 42 , and wherein at least two of the nucleoside subunits are joined by a N3′→P5′ phosphoramidate inter-subunit linkage.47. The method of claim 42 , wherein the polynucleotide comprises the sequence TAGGGTTAGACAA.48. The method of claim 47 , wherein all of the internucleotide inter-subunit linkages of the TAGGGTTAGACAA sequence are N3′→P5′ phosphoramidate inter-subunit linkages.52. The method of claim 47 , wherein the C11 nucleotide residue of the TAGGGTTAGACAA sequence derives from a 3′-protected aminonucleoside-5′-phosphoramidite monomer.53. The method of claim 47 , wherein the method ...

CYCLIC DINUCLEOTIDES FOR TREATING CONDITIONS ASSOCIATED WITH STING ACTIVITY SUCH AS CANCER

This disclosure features dinucleotide compounds that modulate Stimulator of Interferon Genes (STING) activity, for use for example in the treatment of cancer. This disclosure also features compositions as well as other methods of using and making the same (Formula (A)). A and B are each independently selected from the group consisting of Formulae (i), (ii), (iii), and (iv). 1276-. (canceled)292. A compound selected from{'sup': '6,10', '(1S,3R,6S,8R,9R,10S,12R,15S,17R,18R)-8-(2-amino-6-oxo-6,9-dihydro-1H-purin-9-yl)-17-(6-amino-9H-purin-9-yl)-9,18-dihydroxy-3,12-disulfanyl-4,7,13,16-tetraoxa-2,11-diaza-3lambda5,12lambda5-diphosphatricyclo[13.3.0.0]octadecane-3,12-dione;'}{'sup': '6,10', '(1S,3S,6S,8R,9R,10S,12R,15S,17R,18R)-8-(2-amino-6-oxo-6,9-dihydro-1H-purin-9-yl)-17-(6-amino-9H-purin-9-yl)-9,18-dihydroxy-3,12-disulfanyl-4,7,13,16-tetraoxa-2,11-diaza-3lambda5,12lambda5-diphosphatricyclo[13.3.0.0]octadecane-3,12-dione;'}{'sup': '6,10', '(1S,3S,6S,8R,9R,10S,12S,15S,17R,18R)-8-(2-amino-6-oxo-6,9-dihydro-1H-purin-9-yl)-17-(6-amino-9H-purin-9-yl)-9,18-dihydroxy-3,12-disulfanyl-4,7,13,16-tetraoxa-2,11-diaza-3lambda5,12lambda5-diphosphatricyclo[13.3.0.0]octadecane-3,12-dione;'}{'sup': '6,10', '(1S,3R,6S,8R,9R,10S,12S,15S,17R,18R)-8-(2-amino-6-oxo-6,9-dihydro-1H-purin-9-yl)-17-(6-amino-9H-purin-9-yl)-9,18-dihydroxy-3,12-disulfanyl-4,7,13,16-tetraoxa-2,11-diaza-3lambda5,12lambda5-diphosphatricyclo[13.3.0.0]octadecane-3,12-dione; or'}{'sup': '6,10', '(1S,6S,8R,9R,10S,15S,17R,18R)-8-(2-amino-6-oxo-6,9-dihydro-1H-purin-9-yl)-17-(6-amino-9H-purin-9-yl)-3,9,12,18-tetrahydroxy-4,7,13,16-tetraoxa-2,11-diaza-3lambda5,12lambda5-diphosphatricyclo[13.3.0.0]octadecane-3,12-dione.'}293. A pharmaceutical composition comprising a compound according to or a pharmaceutically acceptable salt thereof and one or more pharmaceutically acceptable carriers claim 277 , diluents or excipients.294. A combination pharmaceutical product comprising a compound according to or a pharmaceutically ...

GALNAC PHOSPHORAMIDITES, NUCLEIC ACID CONJUGATES THEREOF AND THEIR USE

This invention generally relates to the field of phosphoramidite derivatives. In particular, the invention relates to N-Acetylgalactosamine phosphoramidite molecules and to conjugates of nucleic acid molecules with N-Acetylgalactosamine containing molecules. Also provided are methods for preparation of these molecules and possible uses thereof, in particular in medicine. 117.-. (canceled)19. Use of a compound according to as a medicament. This application is divisional under 35 U.S.C. § 120 of application Ser. No. 15/517,685 filed Apr. 7, 2017 which is a national stage application under 35 U.S.C. § 371 of PCT Application No. PCT/EP2015/073331 filed Oct. 9, 2015 which claims priority to European Patent Application No. EP14188444.5 filed Oct. 10, 2014 and, European Patent Application No. 15181807.7 filed Aug. 20, 2015, of which each of these applications are hereby incorporated by reference in their entirety.This invention generally relates to the field of phosphoramidite derivatives. In particular, the invention relates to N-Acetylgalactosamine phosphoramidite molecules and to conjugates of nucleic acid molecules with N-Acetylgalactosamine containing molecules. Also provided are methods for preparation of these molecules and possible uses thereof, in particular in medicine.In recent years, approaches have been developed to use nucleic acid molecules in therapy. To favorably influence pharmaceutically relevant properties, the nucleic acid molecules have been conjugated to certain ligands such as peptides, lipids, sterols, and carbohydrates. Nucleic acid conjugates have been extensively evaluated for use in siRNAs, where they are considered essential in order to obtain sufficient in vivo potency. For example, by attachment of a conjugate moiety containing terminal galactose or a derivative thereof to the nucleic acid, thereby targeting the nucleic acid molecule to hepatocytes via binding to the asialoglycoprotein receptor (ASGPR), see for example WO2009/073809, WO2011/ ...

COMPOSITIONS COMPRISING ALTERNATING 2'-MODIFIED NUCLEOSIDES FOR USE IN GENE MODULATION

The present invention provides compositions comprising at least one oligomeric compound comprising an alternating motif and further include a region that is complementary to a nucleic acid target. The compositions are useful for targeting selected nucleic acid molecules and modulating the expression of one or more genes. In preferred embodiments the compositions of the present invention hybridize to a portion of a target RNA resulting in loss of normal function of the target RNA. The present invention also provides methods for modulating gene expression. 1105.-. (canceled)106. A composition comprising a first oligomeric compound and a second oligomeric compound , wherein:at least a portion of said first oligomeric compound is capable of hybridizing with at least a portion of said second oligomeric compound;at least a portion of said first oligomeric compound is complementary to and capable of hybridizing to a selected nucleic acid target;{'sub': 1', '2', '1', '2', '3, 'at least one of said first and said second oligomeric compounds comprises a region having the formula X—Y—X, wherein Y is a region of from about 5 to about 12 linked nucleosides and each of Xand Xis, independently, a plurality of linked nucleosides having the formula FSFS, where one of F and S is a 2′-F modified nucleoside and the other of F and S is a 2′-O—CHmodified nucleoside; and'}each of the linked nucleosides is, independently, linked by a phosphodiester or a phosphorothioate internucleoside linkage.107. The composition of wherein said first oligomeric compound comprises a 5′-phosphate group.108. The composition of wherein said second oligomeric compound comprises a 5′-phosphate group.109. The composition of wherein each of said first and said second oligomeric compounds comprise a 5′-phosphate group.110. The composition of wherein at least one of said first and said second oligomeric compounds comprises at least one conjugate group.111. The composition of wherein at least one of said first and ...

Reagents and methods for post-synthetic modification of nucleic acids

The present invention relates to compositions and methods (reagents and protocols) for the post-synthetic modification of nucleic acids obtained from solid-phase oligonucleotide synthesis with a label (such as fluorescent dyes). The coupling reagent is the triazine-based salt 4-(4,6-dimethoxy-1,3,5-triazin-2-yl)-4-methylmorpholinium (DMT-MM) in the presence of a counteranion.

METHOD FOR DEBLOCKING OF LABELED OLIGONUCLEOTIDES

The invention relates to a process for deblocking substantially a blocked, detectably labeled oligonucleotide by contacting the blocked detectably labeled oligonucleotide with an effective amount of a nucleophilic amino compound under conditions that result in substantial deblocking of the oligonucleotide, thereby giving the substantially deblocked oligonucleotide. 1. A process for deblocking substantially a blocked , detectably labeled oligonucleotide comprising contacting the blocked detectably labeled oligonucleotide with an effective amount of a nucleophilic amino compound under conditions that result in the deblocking of the oligonucleotide , thereby giving the substantially deblocked oligonucleotide.2. The process of claim 1 , wherein said detectable label is a fluorescent label.3. The process of claim 2 , wherein said detectable label is hexachlorofluorescein.4. The process of claim 2 , wherein said detectable label is DABCYL.5. The process of claim 1 , wherein said nucleophilic amino compound is ammonia.6. The process of claim 5 , wherein said ammonia is present at a psi of about 20 to 200.7. The process of claim 5 , wherein said ammonia is present at a psi of about 80.8. The process of claim 1 , wherein said nucleophilic amino compound is ammonia vapors.9. The process of claim 1 , wherein said nucleophilic amino compound is a Calkylamine.10. The process of claim 1 , further comprising dissolving the substantially deblocked oligonucleotide in a buffer.11. The process of claim 1 , wherein said conditions comprise carrying the process at about room temperature to about 150° C.12. The process of claim 1 , wherein said conditions comprise carrying the process at about 95° C.1314-. (canceled)15. The process of claim 1 , wherein said substantially blocked claim 1 , detectably labeled oligonucleotide is immobilized on a solid phase.16. The process of claim 15 , wherein said substantially deblocked oligonucleotide is released from said solid phase under said ...

Cross-Linked Poly-E-Lysine Non-Particulate Support

The invention provides a non-particulate cross-linked poly-ε-lysine polymer. The poly-ε-lysine and cross linker are linked by amide bonds and may the cross linker has at least two functional groups capable of reacting with an alpha carbon amine of poly-ε-lysine. The polymer is suitably insoluble in water and other solvents and is provided in macro form for example a sheet, article or fibre. The macro form polymer is useful in a wide range of applications including wound treatment, as a medical diagnostic comprising a particulate support and a functional material bound or retained by the support and solid phase synthesis of peptides, oligonucleotides, oligosaccharides, immobilisation of species, cell culturing and in chromatographic separation.

FORMYLPYRROLE-BASED HETEROCYCLES FOR NUCLEIC ACID ATTACHMENT TO SUPPORTS

A compound has Formula I: 2. The compound of claim 1 , wherein the protected aldehyde is selected from the group consisting of an acetal claim 1 , an aminal claim 1 , a dithioacetal claim 1 , a protected hemiaminal claim 1 , an alkene claim 1 , and a protected hemithioacetal.3. The compound of claim 1 , wherein W claim 1 , X claim 1 , Y claim 1 , and Z comprise a fused ring system selected from the group consisting of a benzene claim 1 , a pyridine claim 1 , a furan claim 1 , a thiophene claim 1 , a pyridazine claim 1 , a pyrazine claim 1 , and a pyrimidine.4. The compound of claim 3 , wherein W claim 3 , X claim 3 , Y claim 3 , and Z comprise a fused benzene ring.5. The compound of claim 1 , wherein A is hydrogen or methyl claim 1 , Q is a protected aldehyde claim 1 , Ris N-iPr claim 1 , and Ris OCHCHCN.6. The compound of claim 1 , wherein A is a hydroxyl claim 1 , alkoxy or hydroxyalkyl claim 1 , Q is a protected aldehyde claim 1 , Ris N-iPr claim 1 , and Ris OCHCHCN.8. The compound of claim 7 , wherein the protected aldehyde is selected from the group consisting of an acetal claim 7 , an aminal claim 7 , a dithioacetal claim 7 , a protected hemiaminal claim 7 , an alkene claim 7 , and a protected hemithioacetal.9. The compound of wherein W claim 7 , X claim 7 , Y claim 7 , and Z comprise a fused ring system selected from the group consisting of a benzene claim 7 , a pyridine claim 7 , a furan claim 7 , a thiophene claim 7 , a pyridazine claim 7 , a pyrazine claim 7 , and a pyrimidine.10. The compound of claim 9 , wherein W claim 9 , X claim 9 , Y claim 9 , and Z comprise a fused benzene ring.11. The compound of claim 7 , wherein A is hydrogen or methyl claim 7 , Q is a protected aldehyde claim 7 , Nu is selected from the group consisting of a 3′-phosphate-linked nucleic acid claim 7 , a 3′-thiophosphate-linked nucleic acid claim 7 , and a 3′-phosphate linked modified nucleic acid.12. The compound of claim 7 , wherein A is hydrogen or methyl claim 7 , Q is a ...

Enrichment and Sequencing of RNA Species

Provided herein is a method for making an cDNA library, comprising adding an affinity tag-labeled GMP to the 5′ end of targeted RNA species in a sample by optionally decapping followed by incubating the sample with an affinity tag-labeled GTP and a capping enzyme, enriching for RNA comprising the affinity tag-labeled GMP using an affinity matrix that binds to the affinity tag, reverse transcribing the enriched RNA to produce a population of cDNAs, and adding a tail to the 3′ end of the population of cDNAs using a terminal transferase, to produce an cDNA library.

MISPRIMING PREVENTION REAGENTS

Provided herein are mispriming prevention reagents, compositions and kits comprising such reagents and methods of use thereof. 1162-. (canceled)163. A method of creating a cDNA comprising: (i) RNA;', '(iv) a reverse transcriptase;', '(v) dNTPs; and', (1) a first condition-dependent stem region comprising a 5′ terminal covalently linked moiety and a first stem nucleic acid sequence, wherein the first stem nucleic acid sequence is at least 6 nucleotides in length and wherein the 5′ terminal covalently linked moiety comprises a cyclic or polycyclic planar moiety that does not have a bulky portion;', '(2) a condition-dependent loop region comprising a loop nucleic acid sequence of at least 3 nucleotides in length; and', '(3) a second condition-dependent stem region comprising a second stem nucleic acid sequence and a 3′ terminal covalently linked moiety, wherein the second stem nucleic acid sequence is at least 6 nucleotides in length and is complementary to the first stem nucleic acid sequence, wherein the 3′ terminal covalently linked moiety comprises a cyclic or polycyclic planar moiety that does not have a bulky portion, wherein the 3′ terminal covalently linked moiety is non-identical to the 5′ terminal covalently linked moiety, wherein the 3′ terminus of the second condition-dependent stem region is non-extendable by the reverse transcriptase or by a thermostable DNA polymerase, wherein the second condition-dependent stem region hybridizes to the first condition-dependent stem region with a stem melting temperature that is no greater than the first primer melting temperature and the second primer melting temperature, and wherein hybridization of the second condition-dependent stem region to the first condition-dependent stem region causes the reagent to acquire a stem-loop hairpin conformation; and, '(vi) a mispriming prevention reagent comprising a nucleic acid molecule comprising, in 5′ to 3′ order], '(a) forming a reaction mixture comprising(b) incubating the ...

Oligonucleotide Analogues Incorporating 5-Aza-Cytosine Therein

Oligonucleotide analogues are provided that incorporate 5-aza-cytosine in the oligonucleotide sequence, e.g., in the form of 5-aza-2′-deoxycytidine (decitabine) or 5-aza-cytidine. In particular, oligonucleotide analogues rich in decitabine-deoxyguanosine islets (DpG and GpD) are provided to target the CpG islets in the human genome, especially in the promoter regions of genes susceptible to aberrant hypermethylation. Such analogues can be used for modulation of DNA methylation, such as effective inhibition of methylation of cytosine at the C-5 position. Methods for synthesizing these oligonucleotide analogues and for modulating nucleic acid methylation are provided. Also provided are phosphoramidite building blocks for synthesizing the oligonucleotide analogues, methods for synthesizing, formulating and administering these compounds or compositions to treat conditions, such as cancer and hematological disorders. 1114-. (canceled)115. A method of treating a condition , the method comprising administering to a subject in need thereof a therapeutically-effective amount of a dinucleotide analogue , or a pharmaceutically-acceptable salt thereof , wherein the dinucleotide analogue , or the pharmaceutically-acceptable salt thereof , comprises a phospholinker , wherein the number of phosphorus atoms in the phospholinker is one , wherein the linker is not a phosphorothioate linker.116. The method of claim 115 , wherein the dinucleotide analogue or the pharmaceutically-acceptable salt thereof comprises a 5-aza-cytosine group.117. The method of claim 115 , wherein the dinucleotide analogue or the pharmaceutically-acceptable salt thereof comprises a decitabine group.118. The method of claim 115 , wherein the dinucleotide analogue or the pharmaceutically-acceptable salt thereof comprises a deoxyguanosine group.119. The method of claim 115 , wherein the dinucleotide analogue is the pharmaceutically-acceptable salt claim 115 , wherein the pharmaceutically-acceptable salt is a ...

Compositions and method for synthesizing nucleic acids

Provided herein are compositions, kits and methods for synthesis of nucleic acids. Also provided herein are compositions and methods for synthesizing strands of nucleic acid across different nucleic acid back-bones hybridized together using a strand displacing polymerase.

BIOPARTICLE ISOLATION AND THERAPEUTIC APPLICATION THEREOF

Compositions and methods for the isolation of protein-nucleic acid complexes, extracellular vesicle (EV) (e.g., microvesicles) and free nucleic acids (collectively referred to as “bioparticles”) released by mammalian cells into body fluids or cell culture media are provided. Isolated bioparticles of the current disclosure contain biomolecules that are useful as diagnostic/prognostic biomarkers or for identification of therapeutic targets (e.g., disease or disorder-associated miRNAs, circulating tumor DNA). Isolation of biomolecules results in purification and concentration. Methods for producing biofluids without detectable bioparticles, largely depleted of bioparticles, and/or possessing a reduced concentration of bioparticles compared to a biofluid starting material (collectively termed “bioparticle-depleted”) are provided. Bioparticle-depleted biofluid is useful, e.g., in experimental systems where desirable to obtain a biofluid lacking or substantially depleted of endogenous bioparticles from the source material. Non-toxic bioparticle absorbing materials (e.g., exosome-reducing materials) can also be used for prophylactic, therapeutic, validation and/or experimental purposes. 1. A method for isolating , amplifying or both isolating and amplifying cell-free nucleic acids from a liquid sample with enhanced efficiency comprising:a) obtaining a liquid sample from a subject or cell culture;b) contacting said liquid sample with a crystallizing agent under conditions suitable to allow for crystal formation, thereby creating an admixture;c) incubating said admixture for a period of time sufficient to allow for crystal formation;d) separating said admixture to obtain a particle fraction containing bioparticles; ande) isolating amplifying or both isolating and amplifying cell-free nucleic acids from the particle fraction containing bioparticles, thereby isolating, amplifying or both isolating and amplifying cell-free nucleic acids from the liquid sample with enhanced ...

A METHOD FOR THE SITE-SPECIFIC ENZYMATIC LABELLING OF NUCLEIC ACIDS IN VITRO BY INCORPORATION OF UNNATURAL NUCLEOTIDES

Provided herein are analogs of unnatural nucleotides bearing predominantly hydrophobic nucleobase analogs that form unnatural base pairs during DNA polymerase-mediated replication of DNA or RNA polymerase-mediated transcription of RNA. In this manner, the unnatural nucleobases can be introduced in a site-specific way into oligonucleotides (single or double stranded DNA or RNA), where they can provide for site-specific cleavage, or can provide a reactive linker than can undergo functionalization with a cargo-bearing reagent by means of reaction with a primary amino group or by means of click chemistry with an alkyne group of the unnatural nucleobase linker. 120.-. (canceled)22. The method of claim 21 , wherein the method is performed in a cell.23. The method of claim 22 , wherein the cell is a prokaryotic cell.24. The method of claim 21 , wherein Ris hydrogen or halogen.25. The method of claim 24 , wherein the halogen is fluoro.26. The method of claim 24 , wherein Ris hydrogen.27. The method of claim 21 , further comprising retaining the unnatural base pair in the DNA oligonucleotide after two or more rounds of replication.28. The method of claim 27 , wherein retaining is at a rate of at least 90% claim 27 , as measured by polymerase chain reaction (PCR) under standard conditions.29. The method of claim 28 , wherein retaining is at a rate of at least 99% claim 28 , as measured by polymerase chain reaction (PCR) under standard conditions.33. The method of claim 32 , wherein the method is performed in a cell.34. The method of claim 33 , wherein the RNA polymerase is endogenous to the cell.35. The method of claim 33 , wherein the cell is a prokaryotic cell.36. The method of claim 32 , wherein Ris hydrogen or halogen.37. The method of claim 36 , wherein the halogen is fluoro.38. The method of claim 36 , wherein Ris hydrogen. This is a continuation of U.S. patent application Ser. No. 14/910,203, filed Feb. 4, 2016; which is a U.S. National Stage Entry of PCT/US2014/050423 ...

CYCLIC DINUCLEOTIDES CONTAINING BENZIMIDAZOLE, METHOD FOR THE PRODUCTION OF SAME, AND USE OF SAME TO ACTIVATE STIMULATOR OF INTERFERON GENES (STING)-DEPENDENT SIGNALING PATHWAYS

Cyclic dinucleotides are described, which in contrast to their natural congeners carry lipophilic nucleobases and have higher membrane permeability and increased biological activity. 3. The compound according to claim 1 , wherein Rand/or Ris SH; or Rand/or Ris BH; or Ris SH and Ris BH; or Ris BHand Ris SH.4. The compound according to claim 1 , wherein the connections via O* and O** are realized by Rand Rleading to compounds usually assigned as 3′ claim 1 , 5′-3′ claim 1 , 5′-connected.5. The compound according to claim 1 , wherein the connections via O* and O** are realized by Rand Rleading to compounds commonly assigned as 2′ claim 1 , 5′-3′ claim 1 , 5′-connected.6. The compound according to claim 1 , wherein Rand Reach are Cl; or each R claim 1 , Rand Rare Cl; or Ris CFand Rand Reach are Cl.7. The compound according to claim 1 , wherein Ror Rand Ror Reach are OH; or Ror Ris F and Ror Ris OH; or Ror Ris OH and Ror Ris F; or Ror Rand Ror Reach are F.8. The compound according to claim 1 , wherein the structure in accordance with Formula (I) is chosen from:(1) cyclic (benzimidazole riboside-(3′->5′)-monophosphate-benzimidazole riboside-(3′->5′)-monophosphate);(2) cyclic (benzimidazole riboside-(2′->5′)-monophosphate-benzimidazole riboside-(3′->5′)-monophosphate);(3) cyclic (benzimidazole riboside-(2′->5′)-monophosphate-benzimidazole riboside-(2′->5′)-monophosphate);(4) cyclic (benzimidazole riboside-(3′->5′)-monophosphorothioate-benzimidazole riboside-(3′->5′)-monophosphorothioate);(5) cyclic (benzimidazole riboside-(2′->5′)-monophosphorothioate-benzimidazole riboside-(3′->5′)-monophosphorothioate);(6) cyclic (benzimidazole riboside-(2′->5′)-monophosphorothioate-benzimidazole riboside-(2′->5′)-monophosphorothioate);(7) cyclic (5,6-dichlorobenzimidazole riboside-(3′->5′)-monophosphate-5,6-dichlorobenzimi-dazole riboside-(3′->5′)-monophosphate);(8) cyclic (5,6-dichlorobenzimidazole riboside-(2′->5′)-monophosphate-5,6-dichlorobenzimi-dazole riboside-(3′->5′)- ...

COMPOSITIONS AND METHODS FOR MODULATING SMN GENE FAMILY EXPRESSION

Aspects of the invention provide single stranded oligonucleotides for activating or enhancing expression of SMN1 or SMN2. Further aspects provide compositions and kits comprising single stranded oligonucleotides for activating or enhancing expression of SMN1 or SMN2 that comprises exon 7. Methods for modulating expression of SMN1 or SMN2 using the single stranded oligonucleotides are also provided. Further aspects of the invention provide methods for selecting a candidate oligonucleotide for activating or enhancing expression of SMN1 or SMN2. 1. A composition comprising a cell and a single stranded oligonucleotide , wherein the single stranded oligonucleotide is produced by a process comprising:synthesizing a single stranded oligonucleotide that:(a) has a sequence 5′-X-Y-Z, wherein X is any nucleotide, Y is a nucleotide sequence of 6 nucleotides in length that is not a seed sequence of a human microRNA, and Z is a nucleotide sequence of 1-8 nucleotides in length,(b) is 100% complementary with a PRC2-associated region of a human SMN gene, wherein the PRC2-associated region is a region of the SMN gene that has a sequence that occurs at a higher frequency in a sequencing reaction of products of an RNA-immunoprecipitation assay that employs an antibody that targets Ezh2 to immunoprecipitate RNA-associated PRC2 complexes from cells comprising the SMN gene compared to a control sequencing reaction of products of a control RNA-immunoprecipitation assay that employs a control antibody, and(c) is 8 to 15 nucleotides in length,wherein, during the synthesis, at least one nucleotide incorporated into the oligonucleotide comprises a 2′ O-methyl and/or a 2′-fluoro, and a phosphorothioate internucleotide linkage is incorporated between at least two nucleotides.2. The composition of claim 1 , wherein the oligonucleotide does not comprise three or more consecutive guanosine nucleotides.3. The composition of claim 1 , wherein the oligonucleotide does not comprise four or more ...

MODULATORS OF DIACYGLYCEROL ACYLTRANSFERASE 2 (DGAT2)

The present embodiments provide methods, compounds, and compositions useful for inhibiting DGAT2 expression, which may be useful for treating, preventing, or ameliorating a disease associated with DGAT2. 112-. (canceled)13. A compound comprising a modified oligonucleotide consisting of 15 to 30 linked nucleosides , wherein the modified oligonucleotide is 95% complementary within nucleotides 26778-26797 , 23242-23261 , 26630-26649 , 15251-15270 , 28026-28045 , 35436-35455 , 10820-10836 , 23246-23262 of SEQ ID NO: 2.1419-. (canceled)20. The compound of claim 13 , wherein the modified oligonucleotide comprises at least one modified internucleoside linkage claim 13 , at least one modified sugar claim 13 , or at least one modified nucleobase.21. The compound of claim 20 , wherein the modified internucleoside linkage is a phosphorothioate internucleoside linkage.22. The compound of claim 20 , wherein the modified sugar is a bicyclic sugar.23. The compound of claim 22 , wherein the bicyclic sugar is selected from the group consisting of: 4′-(CH)—O-2′ (LNA); 4′-(CH)—O-2′ (ENA); and 4′—CH(CH)—O-2′ (cEt).24. The compound of claim 20 , wherein the modified sugar is 2′—O-methoxyethyl.25. The compound of claim 20 , wherein the modified nucleobase is a 5-methylcytosine.26. The compound of claim 13 , wherein the modified oligonucleotide comprises:a gap segment consisting of linked deoxynucleosides;a 5′ wing segment consisting of linked nucleosides; anda 3′ wing segment consisting of linked nucleosides;wherein the gap segment is positioned immediately adjacent to and between the 5′ wing segment and the 3′ wing segment and wherein each nucleoside of each wing segment comprises a modified sugar.27. The compound of claim 13 , wherein the compound is single-stranded.2833-. (canceled)34. A compound comprising a modified oligonucleotide consisting of 20 linked nucleosides and having a nucleobase sequence consisting of the sequence recited in SEQ ID NO: 1423 claim 13 , wherein the ...

COMPOSITIONS AND METHODS FOR TRANSIENT GENE THERAPY WITH ENHANCED STABILITY

The present invention provides circularized RNA and methods of making, purifying, and using same. 1. A nucleic acid comprising from 5′ to 3′:(a) a 5′ imperfect complement-reverse complement (iCRC) sequence;(b) a 5′ untranslated region (UTR) sequence;(c) an RNA sequence that comprises an open reading frame;(d) a 3′ UTR sequence; and(e) a 3′ iCRC sequence; (i) one or more nucleotide mismatches such that the 5′ iCRC sequence and the 3′ iCRC are not 100% complementary;', {'sub': 'a', '(ii) an annealing temperature (T) above about 16° C.; and'}, {'sub': 'm', '(iii) a melting temperature (T) below about 37° C.'}], 'wherein the 5′ iCRC sequence and the 3′ iCRC sequence have the following characteristics2. A nucleic acid comprising from 5′ to 3′:(a) a 5′ iCRC sequence;(c) a 5′ UTR sequence;(d) an open reading frame;(e) a 3′ UTR sequence; and one or more mismatched base pairs; and', {'sub': 'm', 'a Tof at least 16° C.'}], '(f) a 3′ iCRC sequence, wherein the 5′ iCRC sequence and the 3′ iCRC sequence hybridize under ligation reaction conditions to form a duplex, wherein the duplex comprises3. The nucleic acid according to any one of the preceding claims , further comprising a 5′ tail sequence upstream of the 5′ iCRC sequence and a 3′ tail sequence downstream of the 3′ iCRC sequence , wherein the 5′ tail sequence and the 3′ tail sequence do not hybridize under ligation reaction conditions.4. The nucleic acid of claim 3 , wherein the 5′ tail sequence comprises a G nucleotide at the 5′-most base position.5. The nucleic acid of claim 4 , wherein the 5′ tail sequence comprises between one and three G nucleotides at the 5′-most base positions.6. The nucleic acid according to any one of the preceding claims claim 4 , further comprising at least one polyA sequence.7. The nucleic acid according to any one of the preceding claims claim 4 , wherein a 5′ polyA sequence is located 5′ to the 5′ iCRC sequence and/or a 3′ polyA sequence is located 3′ to the 3′ iCRC sequence.8. The nucleic ...

DEVICE FOR PURIFYING NUCLEIC ACIDS

The invention relates to a device () for purifying nucleic acids composed of a one-piece hollow body () comprising an upper portion () having an inlet port () and a lower portion () having an outlet port (), wherein within the hollow body () at the least one nucleic acid-binding matrix () is arranged, wherein the device () is characterized in that between the upper portion () and the lower portion () a predetermined breaking point () is provided and the nucleic acid-binding matrix () is arranged in the lower portion (). The invention further relates to a method for producing such a device, a method for purifying nucleic acids by means of a device according to the invention, and a kit comprising a device according to the invention. 115-. (canceled)17. The device according to claim 16 , wherein the volume of upper portion corresponds to a volume that is at least 5-fold the volume of lower portion.18. The device according to claim 16 , wherein the upper portion and lower portion independently of one another have a round or rectangular cross section.19. The device according to claim 16 , wherein the upper portion and lower portion independently of one another have a cylindrical or conical form.20. The device according to claim 16 , wherein the hollow body is manufactured of a plastic.21. The device according to claim 16 , wherein the plastic is selected from the groups consisting of polyolefins claim 16 , bio-based plastics claim 16 , polylactates claim 16 , polyamides claim 16 , polyimides claim 16 , acetals claim 16 , polyvinyl chloride claim 16 , polytetrafluoroethylene claim 16 , polyesters claim 16 , polycarbonates claim 16 , polymethyl(meth)acrylates claim 16 , acrylonitrile butatien styrene terpolymer (ABS) claim 16 , polystyrene claim 16 , copolymers thereof claim 16 , and mixtures thereof.22. The device according to claim 16 , wherein the hollow body is manufactured by an injection molding process claim 16 , blow molding claim 16 , casting technique with ...

Oligonucleotide specific uptake of nanoconjugates

Disclosed are nanoparticles functionalized with an oligonucleotide and a domain, wherein the domain increases cellular uptake of the nanoparticles. The domain is a sequence of nucleobases or phosphate groups, such as a poly thymidine (polyT) sequence or a phosphate polymer (C3 residue) and may be located 5′ to the oligonucleotide 3′ to the oligonucleotide, within, or colinear with the oligonucleotide. Usage of the nanoparticles including modulating gene regulation is contemplated.

PROCESS FOR THE PREPARATION OF A CYCLIC DINUCLEOTIDE

The invention generally relates to an improved processes for the preparation of a cyclic dinucleotide which is useful as a STING agonist of the following formula (I), involving the use of compounds A and B. 2. The process according to wherein the base in step b) is imidazole claim 1 , DBU claim 1 , diisopropylethylamine claim 1 , triazole claim 1 , tetrazole or metal alkoxide bases or lithium diisopropylamide claim 1 , sodium bis(trimethysiliyl)amide or other amide bases.3. The process according to wherein the base is imidazole.4. The process according to wherein the solvent in step c) is THF claim 1 , MeCN or DMF.5. The process according to wherein the solvent is THF.6. The process according to wherein the base in step c) is lithium diisopropylamide claim 1 , sodium bis(trimethysiliyl)amide or other amide bases or potassium t-butoxide claim 1 , DBU or other alkoxide bases.7. The process according to wherein the base in step c) is sodium tert-pentoxide or sodium t-amylate.8. The process according to wherein wherein the solvent in step d) is THF claim 1 , 2-MeTHF claim 1 , MeCN or DMF.9. The process according to wherein the solvent in step d) is 2-MeTHF.10. The process according to wherein the base in step d) is imidazole claim 1 , DBU claim 1 , triazole claim 1 , tetrazole claim 1 , diisopropylethylamine or other t-butoxides.11. The process according to wherein the base in step d) is imidazole.12. The process according to wherein the solvent in step f) is THF claim 1 , MeCN claim 1 , NMP or DMF or mixtures thereof.13. The process according to wherein the solvent in step f) is a mixture of THF and NMP.14. The process according to wherein the base in step f) is lithium diisopropylamide claim 1 , potassium bis(trimethylsilyl)amide or other amide bases or potassium t-butoxide claim 1 , DBU or other alkoxide bases or NaHMDS or other alkylsilylamines.15. The process according to wherein the base in step f) is lithium t-butoxide.16. The process according to wherein the ...

DEVICES AND METHODS FOR SYNTHESIS

Provided herein are compositions, devices, systems and methods for electrochemical synthesis. Further provided are devices comprising addressable electrodes controlling polynucleotide synthesis (deprotection, extension, or cleavage, etc.) The compositions, devices, systems and methods described herein provide improved synthesis, storage, density, and retrieval of biomolecule-based information.

Reiterative oligonucleotide synthesis

In some embodiments, the disclosure relates generally to methods, as well as related compositions, systems, and kits, for nucleotide polymerization, oligonucleotide synthesis, detecting nucleotide polymerization, detecting the presence of a nucleic acid, oligonucleotide amplification and detection of oligonucleotide amplification, which can be conducted via an abortive transcription initiation reaction. In some embodiments, abortive transcription initiation reactions can generate multiple copies of an oligonucleotide which can be used to detect the presence of a nucleic acid or macromolecule. In some embodiments, generation of multiple copies of an oligonucleotide can be detected via a sensor that senses the presence of byproducts from a nucleotide incorporation or a nucleotide polymerization reaction. In some embodiments, the byproducts include pyrophosphate, hydrogen ion, charge transfer, and heat. In some embodiments, a abortive transcription initiation reaction can be conducted on a support that can be in contact with or capacitively coupled to at least one sensor. Optionally, the sensor comprises a field-effect transistor (FET).

COMPOUNDS AND METHODS ASSOCIATED WITH DIFFERENTIAL METHYLATION OF HUMAN PAPILLOMA VIRUS GENOMES IN EPITHELIAL CELLS

The invention relates to compounds and methods useful in detection and therapy of HPV-associated diseases. The invention is based on the elucidation of a mechanism by which replication of HPVs occurs in naturally infected tissues and cells. Moreover it is based on the identification of distinct epigenetic changes of the viral genome in infected cells that allows promotion of the affected cells to precancerous and cancerous cells. The invention therefore provides methods of diagnosing neoplasias and their precursor lesions as well as methods of preventing the development of malignancies or inhibiting tumor growth. 1. An isolated nucleic acid comprising a sequence selected from the group consisting of SEQ ID NOs: 1-16.2. A demethylating agent as a medicament for topical application for treatment of an HPV related lesion.3. The demethylating agents of being selected from a group consisting of 5-Azacytidine claim 2 , 5-Aza-20-deoxycytidine claim 2 , Arabinosyl-5-azacytidine claim 2 , 5-6-Dihydro-5-azacytidine claim 2 , 5-Fluoro-20-deoxycytidine claim 2 , Epigallocatechin-3-gallate claim 2 , Hydralazine claim 2 , Procainamide claim 2 , Procaine and Zebularine.4. The demethylating agent of claim 2 , wherein the HPV related lesion is selected from a group consisting of warts claim 2 , exophytic growing papillomas claim 2 , condylomata claim 2 , inverted papillomas claim 2 , pre-neoplastic or neoplastic HPV-induced lesions claim 2 , cervical neoplasias claim 2 , rectal neoplasias claim 2 , neoplasias of the anal epithelium claim 2 , neoplasias of the oropharynx or neoplasias of the tonsils.5. A pharmaceutical composition consisting of a) a demethylating agent and b) a pharmaceutically acceptable carrier.6. The pharmaceutical composition of claim 5 , comprising in addition active ingredients selected from a group comprising buffers claim 5 , humectants claim 5 , excipients claim 5 , preservatives and antiviral substances.7. The pharmaceutical composition of claim 5 , wherein ...

CHEMICAL MODIFICATIONS OF MONOMERS AND OLIGONUCLEOTIDES WITH CYCLOADDITION

The invention features compounds of formula I or II: 3. The oligonucleotide of claim 1 , wherein the ligand is selected from the group consisting of thyrotropin claim 1 , melanotropin claim 1 , lectin claim 1 , glycoprotein claim 1 , surfactant protein A claim 1 , Mucin carbohydrate claim 1 , multivalent lactose claim 1 , multivalent galactose claim 1 , N-acetyl-galactosamine claim 1 , multivalent N-acetyl-galactosamine claim 1 , N-acetyl-glucosamine claim 1 , multivalent N-acetyl glucosamine claim 1 , multivalent mannose claim 1 , multivalent fucose claim 1 , glycosylated polyaminoacids claim 1 , mannose claim 1 , lactose claim 1 , galactose claim 1 , transferrin claim 1 , bisphosphonate claim 1 , polyglutamate claim 1 , polyaspartate claim 1 , a lipid claim 1 , cholesterol claim 1 , a steroid claim 1 , bile acid claim 1 , folate claim 1 , vitamin B12 claim 1 , biotin claim 1 , and an RGD peptide.5. The oligonucleotide of claim 1 , wherein the oligonucleotide is a single-stranded oligonucleotide.6. The oligonucleotide of claim 5 , wherein the single-stranded oligonucleotide is a single-stranded siRNA.7. The oligonucleotide of claim 5 , wherein the single-stranded oligonucleotide is a microRNA.8. The oligonucleotide of claim 1 , wherein the oligonucleotide is a double-stranded oligonucleotide.9. The oligonucleotide of claim 8 , wherein the double-stranded oligonucleotide is a double-stranded siRNA.10. A pharmaceutical composition comprising the oligonucleotide of and a pharmaceutically acceptable excipient.11. The oligonucleotide of claim 1 , wherein Rand Rare H.12. The oligonucleotide of claim 1 , wherein Ris OR.13. The oligonucleotide of claim 1 , wherein Ris OR; and Ris —O-Linker-Q-Linker-R claim 1 , —OC(O)N(H)-Linker-Q-Linker-R claim 1 , or -Linker-Q-Linker-R.16. The oligonucleotide of claim 3 , wherein the ligand is mannose claim 3 , lactose claim 3 , galactose claim 3 , N-acetyl-galactosamine claim 3 , N-acetyl glucosamine claim 3 , multivalent lactose claim 3 ...

POLYMER-BASED EMULSION BREAKING METHODS

A method for breaking emulsions includes applying a polymer mixture to an emulsion. The emulsion can be energized, such as through centrifugation or vibration. In particular, the polymer mixture can be in liquid form. The polymer mixture includes first and second liquid polymer, the second liquid polymer being less hydrophilic than the first liquid polymer. In a water-in-oil emulsion, the less hydrophilic polymer can preferentially reside within the oil phase. 1. A method of recovering a particle from an emulsion , the method comprising:contacting the emulsion with a breaking solution, the emulsion comprising an aqueous phase dispersed in an immiscible continuous phase, the aqueous phase including hydrophilic particles, the breaking solution comprising a first liquid polymer and a second liquid polymer, the first liquid polymer having affinity for the aqueous phase and the second liquid polymer having affinity for the immiscible continuous phase, the molecular weight of the first liquid polymer being less than the molecular weight of the second liquid polymer, wherein the contacting breaks the emulsion, providing a continuous aqueous phase including the hydrophilic particles; andseparating the immiscible continuous phase from the continuous aqueous phase.2. The method of claim 1 , wherein the breaking solution has a total polymer content in a range of 10% to 100% by volume.3. The method of claim 1 , wherein the breaking solution includes the first liquid polymer and the second liquid polymer in a ratio of 1:3 to 10:1 (second liquid polymer:first liquid polymer).4. The method of claim 1 , wherein the first liquid polymer has a molecular weight in a range of 50 Da to 500 Da.5. The method of claim 1 , wherein the second liquid polymer has a molecular weight in a range of 100 Da to 700 Da.6. The method of claim 1 , wherein the emulsion is contact with the breaking solution in a ratio in a range of 20:1 to 1:10.7. The method of claim 1 , wherein the first liquid polymer ...

IONIC LIQUID SUPPORTED SYNTHESIS

The present invention relates to ionic liquids for use in chemical applications and capable of serving the dual function of solvent and liquid support. The ionic liquid lends itself to a method of synthesizing oligomers selected from the group consisting of oligopeptides, oligosaccharides and oligonucleotides, comprising contacting a first monomer unit with an ionic liquid at reaction conditions to provide an ionic liquid bound monomer unit; and contacting the ionic liquid bound monomer unit with at least one further monomer unit at reaction conditions to provide an ionic liquid bound oligomer comprising from 2 to 30 monomer units. The method lends itself to large scale manufacture of oligopeptides, oligosaccharides and oligonucleotides. 131-. (canceled)32. A kit comprising at least one ionic liquid , wherein the ionic liquid is an organic salt comprising:a heterocyclic or substituted heterocyclic quaternary nitrogen-containing organic cation, a heterocyclic or substituted heterocyclic quaternary phosphonium containing organic cation, or a heterocyclic or substituted heterocyclic trivalent sulfonium containing organic cation, andan anion balancing the charge on the organic cation,and monomeric units selected from amino acids, saccharides and nucleotides.33. The kit of claim 32 , suitable for producing an oligopeptide claim 32 , an oligosaccharide or an oligonucleotide comprising at least 2 monomeric units.34. An article of manufacture comprising at least one ionic liquid claim 32 , wherein the ionic liquid is an organic salt comprising:a heterocyclic or substituted heterocyclic quaternary nitrogen-containing organic cation, a heterocyclic or substituted heterocyclic quaternary phosphonium containing organic cation, or a heterocyclic or substituted heterocyclic trivalent sulfonium containing organic cation, andan anion balancing the charge on the organic cation,and further comprising monomeric units selected from amino acids, saccharides and nucleotides.35. The ...

MATERIAL FOR SUPPORTED SYNTHESIS AND METHOD FOR GROWING OLIGONUCLEOTIDES OR PEPTIDES

The invention concerns a material composed of a porous support on which functionalized nanoparticles are grafted by covalent bonding, characterized in that at least part of the nanoparticles grafted by covalent bonding is housed inside surface pores of the support, and in that the support is silica-based and is in the form of porous particles of heterogeneous shape and size, the size of the particles being larger than 1 μm and preferably within the range of 5 to 200 μm. 1- A material composed of a porous support on which functionalized nanoparticles are grafted by covalent bonding , characterized in that at least a portion of the nanoparticles grafted by covalent bonding is housed inside surface pores of the support , and in that the support is silica-based and is in the form of porous particles of heterogeneous shape and size , the size of the particles being larger than 1 μm , and preferably within the range of 5 to 200 μm.2- The material according to claim 1 , characterized that the support is a mineral glass containing silica or a silicate.3- The material according to claim 1 , characterized in that the support is composed exclusively of silica or is composed exclusively of a silicate in which the silica is in a mixture with another or other oxides such as BOto form borosilicates or AlOto formal uminosilicates claim 1 , optionally in a mixture with NaO.4- The material according to claim 1 , characterized in that the minimum mean size of the surface openings formed by the surface pores of the support is at least 3 times equal to the size of the nanoparticles.5- The material according to claim 1 , characterized in that the mean diameter of the pores of the support is in the range of 30 to 600 nm claim 1 , preferably in the range of 100 to 400 nm.6- The material according to claim 1 , characterized in that the support has homogeneous porosity corresponding to the fact that at least 80% of the pores have a diameter whose value does not vary by more than 10% compared ...

DOUBLE COUPLING METHOD FOR OLIGONUCLEOTIDE SYNTHESIS

Aspects of the present disclosure include methods for double coupling a nucleoside phosphoramidite during synthesis of an oligonucleotide. The method can include coupling a free hydroxyl group of a nucleoside residue with a first sample of a protected nucleoside phosphoramidite via an internucleoside P(III) linkage, followed by exposure to an oxidizing agent prior to a second coupling step with a second sample of the protected nucleoside phosphoramidite, and further exposure to an oxidizing agent. The method finds use in synthesizing an oligonucleotide on a solid phase support, such as a planar surface. The double coupling method can be utilized at one or more nucleotide positions during oligonucleotide synthesis thereby reducing single base deletion rates. Oligonucleotide containing compositions synthesized according to the disclosed methods are also provided. 1. A method for double coupling a nucleoside phosphoramidite during synthesis of an oligonucleotide , the method comprising:(a) contacting a free hydroxyl group of a terminal nucleoside residue attached to a solid phase support with a first sample of a protected nucleoside phosphoramidite to couple the protected nucleoside to the terminal nucleoside residue via an internucleoside P(III) linkage;(b) exposing the contacted nucleoside residue to an oxidizing agent to oxidize the linkage and produce a first coupled and oxidized product;(c) contacting the first coupled and oxidized product with a second sample of the protected nucleoside phosphoramidite to couple the protected nucleoside to residual free hydroxyl groups of the terminal nucleoside residue via an internucleoside P(III) linkage; and(d) after step (c), adding an oxidizing agent to oxidize the linkage and produce a protected terminal nucleoside residue.2. The method of claim 1 , further comprising:(e) deprotecting the protected hydroxyl group of the terminal nucleoside residue to produce a free hydroxyl group;repeating steps (a) through (e) until the ...

Multimeric polynucleotides and uses thereof

Aspects of the disclosure relate to multimeric molecules and methods of producing the same. In some embodiments, the multimeric molecules comprise at least two nucleic acid molecules (e.g., mRNA molecules) joined by covalent bonds between non-coding regions.

OLIGONUCLEOTIDE SYNTHESIZER

The disclosure provides a new and improved oligonucleotide synthesizer. 1. A process for making an oligonucleotide , comprising reacting a oligonucleotide precursor with a solid phase support , and mechanically stirring the solid phase support.2. The process of claim 1 , further comprising one or more activation steps.3. The process of claim 1 , further comprising one or more coupling steps.4. The process of claim 1 , further comprising one or more capping steps.5. The process of claim 1 , further comprising one or more oxidation steps.6. The process of claim 1 , further comprising one or more detritylation steps.7. The process of claim 1 , further comprising one or more cleavage steps.8. The process of claim 1 , further comprising one or more deprotection steps.9. The process of claim 1 , wherein the solid phase support comprises a plurality of discrete resin pieces.10. The process of claim 9 , wherein the plurality of discrete resin pieces comprises a plurality of resin beads.11. The process of claim 1 , wherein the mechanical stirring of the solid phase support comprises the motion of a reaction vessel comprising the solid support relative to a chassis claim 1 , frame claim 1 , or support.12. The process of claim 11 , wherein the motion comprises one or more of tipping claim 11 , rocking claim 11 , shaking claim 11 , and inverting the reaction vessel relative to the chassis claim 11 , frame claim 11 , or support.13. The process of claim 1 , wherein the mechanical stirring of the solid phase support comprises stirring via a stir blade claim 1 , a stir bar claim 1 , or both.14. The process of claim 1 , wherein the mechanical stirring of the solid phase support comprises stirring via a plurality of bubbles moving relative to the solid phase support.15. The process of claim 9 , wherein the plurality of discrete resin pieces comprise a low cross link resin for high swelling in acetonitrile (ACN) claim 9 , wherein the low cross link resin has a higher substitution ...

COMPOSITIONS AND METHODS FOR MODULATING PKK EXPRESSION

Disclosed herein are antisense compounds and methods for decreasing PKK mRNA and protein expression. Such methods, compounds, and compositions are useful to treat, prevent, or ameliorate PKK-associated diseases, disorders, and conditions. 1219.-. (canceled)220. A compound comprising a modified oligonucleotide and a conjugate group , wherein the modified oligonucleotide consists of 12 to 30 linked nucleosides and has a nucleobase sequence comprising a portion of at least 15 contiguous nucleobases that is at least 90% complementary to an equal length portion of nucleobases 33183-33242 of SEQ ID NO: 10 , and wherein the conjugate group comprises at least one N-Acetylgalactosamine (GalNAc).221. The compound of claim 220 , wherein the portion of at least 15 contiguous nucleobases is 100% complementary to the equal length portion of nucleobases 33183-33242 of SEQ ID NO: 10.222. The compound of claim 220 , wherein the sequence of the modified oligonucleotide is SEQ ID NO: 705.223. The compound of claim 220 , wherein the conjugate group comprises three GalNAcs.225. The compound of claim 224 , consisting of the modified oligonucleotide and the conjugate group.226. The compound of claim 220 , wherein the modified oligonucleotide consists of 20 linked nucleosides.227. The compound of claim 220 , wherein the modified oligonucleotide is at least 90% complementary to SEQ ID NO: 10.228. The compound of claim 220 , wherein at least one internucleoside linkage of the modified oligonucleotide is a phosphorothioate linkage.229. The compound of claim 220 , wherein each cytosine of the modified oligonucleotide is a 5′-methylcytosine.230. The compound of claim 220 , wherein the modified oligonucleotide is single-stranded.231. The compound of claim 220 , comprising at least one 2′-O-methoxyethyl nucleoside claim 220 , 2′-O-methyl nucleoside claim 220 , constrained ethyl nucleoside claim 220 , LNA nucleoside claim 220 , or 3′-fluoro-HNA nucleoside.232. The compound of claim 220 , wherein ...

CROSS-LINKED POLY-E-LYSINE NON-PARTICULATE SUPPORT

The invention provides a non-particulate cross-linked poly-ε-lysine polymer. The poly-ε-lysine and cross linker are linked by amide bonds and may the cross linker has at least two functional groups capable of reacting with an alpha carbon amine of poly-ε-lysine. The polymer is suitably insoluble in water and other solvents and is provided in macro form for example a sheet, article or fibre. The macro form polymer is useful in a wide range of applications including wound treatment, as a medical diagnostic comprising a particulate support and a functional material bound or retained by the support and solid phase synthesis of peptides, oligonucleotides, oligosaccharides, immobilisation of species, cell culturing and in chromatographic separation. 1. A non-particulate cross-linked poly-ε-lysine polymer comprising poly-ε-lysine and a cross linker linked by amide bonds wherein the cross linker comprises at least two functional groups capable of reacting with an alpha carbon amine of poly-ε-lysine.2. A polymer according to wherein the cross-linker comprises a moiety derived from a compound of formula X[COH]where n is 2 or more and X is a hydrophobic or hydrophilic linking group having a molecular weight of 14 to 250 excluding any functional substituents on the linking group.3. A polymer according to which is insoluble in water.4. A polymer according to which is porous.5. A non-particulate support comprising a polymer according to .6. A non-particulate support according to wherein the cross-linked poly-ε-lysine support is used to coat a non-particulate media directly or indirectly.7. A non-particulate support according to wherein the cross-linked poly-ε-lysine support is used to coat and is bound covalently to a non-particulate media.8. A non-particulate support according to wherein the cross-linked poly-ε-lysine is used to coat an organic non-particulate media.9. A non-particulate support according to wherein the cross-linked poly-ε-lysine is used to coat an inorganic non- ...

Dithiolane Based Thiol Modifier For Labeling and Stronger Immobilization of Bio-Molecules On Solid Surfaces

The thiol modified oligonucleotides have vast number of applications in the field of nucleic acid chemistry. The conjugates generated by mono thiol groups are unstable at higher temperature, in high salt concentration buffers and in presence other thiols. There is strong need to develop a novel thiol modifier probes that can generate multiple thiol groups. Described herein are efficient processes and compounds, dithiolane phosphoramidites derivative and dithiolane succinyl supports. The advantage of our cyclic disulfide thiol modifier is multifold a) each incorporation introduces two thiol groups; b) it can be introduced at any desired site of oligonucleotides; c) The symmetrical branching nature of the spacer in the linker arm of dithiolane allows for clean oligo synthesis, where cleavage of the linker arm and thereby of loss of oligo chain is prevented. We have successfully made 20-mer oligonucleotide containing single dithiolane derivative at 3′, and 21-mer oligonucleotides containing single dithiolane derivative at 5′ or in the middle of the mixed base sequence. HPLC and ESI MS analysis of these oligonucleotides indicated satisfactory purity and correct composition of these oligos, respectively.

COMPOSITIONS AND METHODS FOR ALTERING SECOND MESSENGER SIGNALING

The invention relates to compositions, methods, kits, and assays related to the use and/or exploitation of isomers of cGAMP as well as the structure of the enzyme cGAS. 1. A modulator of cGAS having a structure comprising the following features:{'br': None, 'A-L-B'}wherein:A is or comprises a moiety that fits in the cGAS adenosine binding site;B is or comprises a moiety that fits in the cGAS guanosine binding site; andL is a linker moiety linking A and B in a manner which allows A and B to adopt appropriate interactions to bind cGAS.9. The compound of any one of the preceding claims , wherein each Xis —N—.10. The compound of any one of preceding claims , wherein each Xis —N—.11. The compound of any one of the preceding claims , wherein Xis —NR—.12. The compound of any one of the preceding claims , wherein Xis —O—.13. The compound of any one of the preceding claims , wherein Xis —O—.14. The compound of any one of the preceding claims , wherein Xand Xare —C(R)—.15. The compound of any one of the preceding claims , wherein Xand Xare both oxygen.16. The compound of any one of the preceding claims , wherein Xand Xare both oxygen.17. The compound of any one of the preceding claims , wherein Ris selected from the group consisting of hydrogen , halogen , —OR , —SR , —N(R) , and optionally substituted Caliphatic or Calkoxy-Calkyl.18. The compound of claim 17 , wherein Ris —OH.19. The compound of any one of the preceding claims claim 17 , wherein Ris selected from the group consisting of hydrogen claim 17 , halogen claim 17 , —OR claim 17 , —SR claim 17 , —N(R) claim 17 , and optionally substituted Caliphatic or Calkoxy-Calkyl.20. The compound of claim 19 , wherein Ris —OH.21. The compound of or claim 19 , wherein Ris R.22. The compound of any one of the preceding claims claim 19 , wherein each R claim 19 , R claim 19 , and Ris independently selected from the group consisting of hydrogen claim 19 , halogen claim 19 , and optionally substituted Caliphatic.23. The compound of ...

CYCLIC DI-NUCLEOTIDE COMPOUNDS AS STING AGONISTS

A class of polycyclic compounds of general formula (I), wherein Base, Base, Y, Y, X, X, X, X, X, X, X, X, R, R, R, R, R, R, R, R, R, R, R, R, R, R, and Rare defined herein, that may be useful as inductors of type I interferon production, specifically as STING active agents, are provided. Also provided are processes for the synthesis and use of compounds. 10. A pharmaceutical composition , said pharmaceutical composition comprising:{'claim-ref': {'@idref': 'CLM-00001', 'claim 1'}, '(a) a compound according to or a pharmaceutically acceptable salt thereof; and'}(b) a pharmaceutically acceptable carrier.11. A method of inducing an immune response in a subject claim 1 , said method comprising administering a therapeutically effective amount of a compound according to to the subject.12. A method of inducing an immune response in a subject claim 10 , said method comprising administering a therapeutically effective amount of a pharmaceutical composition according to to the subject.13. A method of inducing a STING-dependent type I interferon production in a subject claim 1 , said method comprising administering a therapeutically effective amount of a compound according to to the subject.14. A method of inducing a STING-dependent type I interferon production in a subject claim 10 , said method comprising administering a therapeutically effective amount of a pharmaceutical composition according to to the subject.15. A method of treating a cell proliferation disorder in a subject claim 1 , said method comprising administering a therapeutically effective amount of a compound according to to the subject.16. The method of claim 15 , wherein the cell proliferation disorder is cancer.17. A method of treating a cell proliferation disorder in a subject claim 10 , said method comprising administering a therapeutically effective amount of a pharmaceutical composition according to to the subject.18. The method of claim 17 , wherein the cell proliferation disorder is cancer. The present ...

RAPID AND EFFICIENT BIOORTHOGONAL LIGATION REACTION AND BORON-CONTAINING HETEROCYCLES USEFUL IN CONJUNCTION THEREWITH

A reaction method comprising combining a carbonyl-substituted arylboronic acid or ester and an α-effect amine in aqueous solution at a temperature between about −5 C to 55 C, and a pH between 2 and 8 to produce an adduct. A process is also provided comprising: contacting a boron compound having a boron atom bonded to a sphybridized carbon conjugated with a cis-carbonyl, the boron having at least one labile substituent, with an α-effect amine, in a solvent for a time sufficient to form an adduct, which may proceed to further products. 1. A process comprising: [{'sup': '2', '(1) a boron compound having a boron atom covalently bonded to an sphybridized carbon conjugated with a cis-carbonyl; and'}, '(2) an α-effect amine; and, '(a) providing(b) contacting the boron compound with the α-effect amine in a liquid solvent at a temperature of less than 55° C. and at a pH above 2, to spontaneously form an adduct by formation of a covalent bond between the boron compound and the α-effect amine.2. The process according to claim 1 , wherein the solvent comprises an aqueous medium claim 1 , the contacting being performed at temperatures between about −5° C. to 55° C. claim 1 , and at a pH between 2 and 8 claim 1 , and wherein the boron compound claim 1 , the α-effect amine claim 1 , and the solvent are bioorthogonal.3. The process according to claim 1 , wherein the α-effect amine comprises an amine group bonded to an oxygen which is reactive with the boron compound.4. The process according to claim 1 , wherein the α-effect amine comprises an amine group bonded to a nitrogen which is reactive with the boron compound.5. The process according to claim 1 , wherein the boron compound comprises a carbonyl-substituted arylboronic acid or ester.7. The process according to claim 6 , wherein:{'sub': 2', '3, 'Ris selected from the group consisting of: H, CH; and'}{'sub': '3', 'Ris selected from the group consisting of OH, O-alkyl, O-alkylbromide, O-alkylamine, O-alkylamide, O-alkylthiol, O- ...

Antisense-induced exon exclusion in myostatin

The present disclosure relates to antisense oligomers and related compositions and methods for decreasing the expression of functional human myostatin and methods for treating muscular dystrophy and related disorders and more specifically relates to inducing exclusion of myostatin exon 2 and thereby reducing levels of myostatin protein.

Oligonucleotide with protected base

The present invention provides a protected nucleotide for elongation, which can be purified efficiently and in a high yield by a liquid-liquid extraction operation, and can achieve an oligonucleotide production method by a phosphoramidite method. It has been found that the above-mentioned problem can be solved by a particular oligonucleotide comprising a protected base and/or particular oligonucleotide protected by a branched chain-containing aromatic group at 3′-position.

METHOD AND KIT FOR CONCENTRATING TARGET DOUBLE-STRANDED NUCLEIC ACID MOLECULES USING A PYRROLE-IMIDAZOLE-CONTAINING POLYAMIDE

A method of separating a target double-stranded nucleic acid molecule from a sample including the target double-stranded nucleic acid molecule and a non-target double-stranded nucleic acid molecule, including (1) mixing the sample, a pyrrole-imidazole-containing polyamide (first PI polyamide) modified with a first linker molecule and capable of specifically binding to a sequence of the target double-stranded nucleic acid molecule, and a carrier a modified with a first ligand capable of specifically binding and/or adsorbing to the first linker molecule such that a mixed solution is produced, (2) forming a complex A by binding the carrier a to the first PI polyamide with which the target double-stranded nucleic acid molecule is bound in the mixed solution, and (3) separating the complex A from the mixed solution. 1. A method of separating a target double-stranded nucleic acid molecule from a sample , comprising:mixing a sample, a first PI polyamide, and a carrier such that a mixed solution comprising the sample, the first PI polyamide, and the carrier is produced;forming a complex A by binding the carrier to the first PI polyamide with which a target double-stranded nucleic acid molecule in the sample is bound in the mixed solution; andseparating the complex A from the mixed solution,wherein the sample includes the target double-stranded nucleic acid molecule and a non-target double-stranded nucleic acid molecule, the first PI polyamide is a pyrrole-imidazole-containing polyamide which is modified with a first linker molecule, includes a plurality of β-alanine moieties such that the pyrrole-imidazole-containing polyamide has at least one β-alanine moiety in every two residues or three residues in a sequence of the pyrrole-imidazole-containing polyamide, and specifically binds to a sequence of the target double-stranded nucleic acid molecule in the sample, and the carrier is modified with a first ligand which specifically binds or adsorbs to the first linker molecule.2 ...

METHODS OF SYNTHESIZING LABELED NUCLEOSIDES

Disclosed herein, inter alia, are compounds, compositions, and methods of synthesizing labeled nucleosides. 2. The method of claim 1 , wherein the methylthiomethyl donor is dimethyl sulfoxide (DMSO).4. The method of claim 3 , wherein Ris substituted or unsubstituted alkyl.5. The method of claim 3 , wherein Ris substituted or unsubstituted C-Calkyl.6. The method of claim 3 , wherein Ris unsubstituted C-Calkyl.7. The method of claim 3 , wherein Ris methyl claim 3 , ethyl claim 3 , isopropyl claim 3 , n-propyl claim 3 , n-butyl claim 3 , sec-butyl claim 3 , isobutyl claim 3 , or tert-butyl.8. The method of claim 3 , wherein Ris tert-butyl.9. The method of claim 1 , wherein B is a protected nucleobase.10. The method of claim 1 , wherein B is a protected nucleobase substituted with a covalent linker to a reactive group.11. The method of claim 1 , wherein B is a substituted or unsubstituted cytosinyl claim 1 , substituted or unsubstituted guaninyl claim 1 , substituted or unsubstituted adeninyl claim 1 , substituted or unsubstituted thyminyl claim 1 , substituted or unsubstituted uracilyl claim 1 , substituted or unsubstituted hypoxanthinyl claim 1 , substituted or unsubstituted xanthinyl claim 1 , substituted or unsubstituted deaza-adeninyl claim 1 , substituted or unsubstituted deaza-guaninyl claim 1 , substituted or unsubstituted deaza-hypoxanthinyl claim 1 , substituted or unsubstituted 7-methylguaninyl claim 1 , substituted or unsubstituted 5 claim 1 ,6-dihydrouracilyl claim 1 , substituted or unsubstituted 5-methylcytosinyl claim 1 , or substituted or unsubstituted 5-hydroxymethylcytosinyl. This application claims the benefit of U.S. Provisional Application No. 62/558,181, filed Sep. 13, 2017, which is incorporated herein by reference in its entirety and for all purposes.DNA sequencing is a fundamental tool in biological and medical research, and is especially important for the paradigm of personalized medicine. Various new DNA sequencing methods have been ...

POLYMERASE-MEDIATED, TEMPLATE-INDEPENDENT POLYNUCLEOTIDE SYNTHESIS

Methods for de novo synthesis of polynucleotides in which 3′-O-reversibly blocked nucleotides are attached to a solid support in the presence of an X family DNA polymerase and in the absence of a nucleic acid template. 1. A method for synthesizing a polynucleotide , wherein the method is tem plate-independent and initiator sequence-independent , and the method comprises:(a) providing a solid support comprising a free hydroxyl group, wherein the free hydroxyl group is part of a cleavable group linked to the solid support;(b) contacting the free hydroxyl group with a nucleotide 5′-triphosphate comprising a removable 3′-O-blocking group in the presence of an X family DNA polymerase and absence of a nucleic acid template to form an immobilized nucleotide comprising a removable 3′-O-blocking group;(c) contacting the immobilized nucleotide comprising the removable 3′-O-blocking group with a deblocking agent to remove the removable 3′-O-blocking group;(d) repeating steps (b) and (c) to yield the polynucleotide; and(e) cleaving the cleavable group of the solid support to release the polynucleotide.2. The method of claim 1 , wherein the cleavable group is attached to the solid support via a linker.3. The method of claim 1 , wherein the cleavable group is a photocleavable group.4. The method of claim 1 , wherein the solid support is a polymer chosen from polypropylene claim 1 , polyethylene claim 1 , cyclo-olefin polymer claim 1 , or cyclo-olefin copolymer.5. The method of claim 1 , wherein the nucleotide 5′-triphosphate comprising the removable 3′-O-blocking group has a sugar moiety chosen from ribose claim 1 , 2′-deoxyribose claim 1 , or 2′-4′ locked deoxyribose and a nitrogenous base chosen from a standard nucleobase claim 1 , a non-standard base claim 1 , a modified base claim 1 , an artificial base claim 1 , or an analog thereof.6. The method of claim 5 , wherein the removable 3′-O-blocking group is chosen from (CO)R claim 5 , (CO)OR claim 5 , (CO)CHOR claim 5 , (CO)NHR ...

MICROFLUIDIC REACTORS FOR OLIGONUCLEOTIDE SYNTHESIS

The present disclosure generally pertains to systems and methods for the chemical synthesis of micro-quantities of oligonucleotides or other chemical molecules. The system includes a reusable glass micro-reactor containing a paramagnetic solid support, a magnet, an electronic drive controller and an optical spectroscopy system capable of driving a plurality individual reactors. The system utilizes the electroosmotic movement of reactants through microfluidic channels. Spectrophotometric monitoring of the reaction products allows for the real-time determination of synthesis yield. 1. An apparatus for synthesizing a molecule , comprising:a reaction substrate having at least one reaction chamber and at least one microfluidic channel for transferring a reagent to the reaction chamber;a reagent cassette coupled to the substrate, the reagent cassette having at least one reagent port for transferring the reagent from the cassette to the microfluidic channel;a paramagnetic solid support positioned within the reaction chamber; anda magnet positioned such that the paramagnetic solid support is located within a magnetic field generated in the magnet, wherein the magnetic field is sufficient for holding the paramagnetic solid support in the reaction chamber while the reagent is flowing through the reaction chamber.2. The apparatus of claim 1 , further comprising a power supply configured to apply a voltage differential claim 1 , wherein the voltage differential is sufficient for generating an electroosmotic force to cause the reagent to flow from the cassette through the microfluidic channel.3. The apparatus of claim 1 , further comprising a spectrophotometer positioned near the microfluidic channel.4. The apparatus of claim 1 , wherein the reaction substrate comprises photodefinable glass.5. The apparatus of claim 1 , further comprising a plurality of reaction substrates.6. The apparatus of claim 1 , wherein the reaction substrate comprises a plurality of reaction chambers.7. ...

Process for galnac oligonucleotide conjugates

The invention comprises a process for the preparation of therapeutically valuable GalNAc cluster oligonucleotide conjugates. The process comprises the coupling of an alkali metal salt, earth alkali metal salt or a tetraalkylammonium salt of an oligonucleotide with a GalNAc cluster compound or with a salt thereof and a subsequent purification.

OLIGONUCLEOTIDES COMPRISING ACYCLIC AND ABASIC NUCLEOSIDES AND ANALOGS

This invention relates to an oligonucleotide comprising one or more abasic nucleoside monomers of formula IV′: 2. The monomer of claim 1 , wherein Ris a protecting group and Ris a reactive phosphorous group or solid support.3. The monomer of claim 1 , wherein Ris a reactive phosphorous group or solid support and Ris a protecting group.4. The monomer of claim 1 , wherein the reactive phosphorus group is selected from the group consisting of phosphoramidite claim 1 , H-phosphonate claim 1 , alkyl-phosphonate claim 1 , and phosphate triester.5. The monomer of claim 1 , wherein the protecting group is a hydroxyl protecting group selected from the group consisting of acetyl claim 1 , benzyl claim 1 , t-butyldimethylsilyl claim 1 , t-butyldiphenylsilyl claim 1 , trityl claim 1 , monomethoxytrityl claim 1 , and dimethoxytrityl.6. An oligonucleotide comprising at least one monomer of .9. The oligonucleotide of claim 7 , wherein the monomer is at the 5′-end terminal position of the oligonucleotide.10. The oligonucleotide of claim 7 , wherein the monomer is at the 3′-end terminal position of the oligonucleotide.11. The oligonucleotide of claim 7 , wherein the monomer is at an internal position of the oligonucleotide.12. The oligonucleotide of claim 7 , wherein the oligonucleotide comprises at least one non-phosphodiester internucleoside linkage.13. The oligonucleotide of claim 7 , wherein the oligonucleotide comprises at least one non-phosphodiester internucleoside linkage selected from the group consisting of phosphorothioate claim 7 , phosphorodithioate claim 7 , H-phosphonate claim 7 , alkyl-phosphonate claim 7 , phosphoramidate internucleoside linkages claim 7 , and any combinations thereof.14. The oligonucleotide of claim 7 , wherein the oligonucleotide comprises at least one nucleobase modification.15. The oligonucleotide of claim 7 , wherein the oligonucleotide comprises at least one sugar modification.16. The oligonucleotide of claim 7 , wherein the oligonucleotide ...

NOVEL LIPID FORMULATIONS FOR NUCLEIC ACID DELIVERY

The present invention provides novel, stable lipid particles comprising one or more active agents or therapeutic agents, methods of making the lipid particles, and methods of delivering and/or administering the lipid particles. More particularly, the present invention provides stable nucleic acid-lipid particles (SNALP) comprising a nucleic acid (such as one or more interfering RNA), methods of making the SNALP, and methods of delivering and/or administering the SNALP. 1. A nucleic acid-lipid particle comprising:(a) a nucleic acid;(b) a cationic lipid comprising from about 50 mol % to about 85 mol % of the total lipid present in the particle;(c) a non-cationic lipid comprising from about 13 mol % to about 49.5 mol % of the total lipid present in the particle; and(d) a conjugated lipid that inhibits aggregation of particles comprising from about 0.5 mol % to about 2 mol % of the total lipid present in the particle.2. The nucleic acid-lipid particle of claim 1 , wherein the nucleic acid comprises a small interfering RNA (siRNA).3. The nucleic acid-lipid particle of claim 2 , wherein the siRNA comprises from about 15 to about 60 nucleotides.4. The nucleic acid-lipid particle of claim 2 , wherein the siRNA comprises at least one modified nucleotide.5. The nucleic acid-lipid particle of claim 2 , wherein the siRNA comprises at least one 2′-O-methyl (2′OMe) nucleotide.6. The nucleic acid-lipid particle of claim 1 , wherein the cationic lipid comprises 1 claim 1 ,2-dilinoleyloxy-N claim 1 ,N-dimethylaminopropane (DLinDMA) claim 1 , 1 claim 1 ,2-dilinolenyloxy-N claim 1 ,N-dimethylaminopropane (DLenDMA) claim 1 , or a mixture thereof.7. The nucleic acid-lipid particle of claim 1 , wherein the cationic lipid comprises 2 claim 1 ,2-dilinoleyl-4-(2-dimethylaminoethyl)-[1 claim 1 ,3]-dioxolane (DLin-K-C2-DMA).8. The nucleic acid-lipid particle of claim 1 , wherein the cationic lipid comprises from about 56.5 mol % to about 66.5 mol % of the total lipid present in the particle.9 ...

Methods of inserting molecular barcodes

The present invention provides compositions, methods, and kits for inserting a plurality of synthetic transposons each comprising a different nucleic acid sequence (i.e., molecular barcode) in a target nucleic acid of interest to allow extraction of contiguity information in the target nucleic acid. The molecular barcodes are also useful for reducing amplification or sequencing bias and errors, and for guiding accurate sequence assembly of the target nucleic acid from sequencing reads. The compositions, methods, and kits described herein have many applications, including haplotyping, genome assembly, sequencing of repetitive regions, detection of structural variations and copy number variations, chromosomal conformation analysis, and methylation analysis.

MAGNETICALLY ASSISTED PROCESSING OF A MEDIUM

The invention relates to a processing device () and a method for processing a medium in a processing chamber (). The processing comprises the addition of magnetic particles (M) to the medium and the mixing of the medium by manipulating said magnetic particles with a time-variable magnetic field (B), particularly a partially oscillating or rotating field. The magnetic field (B) may be generated with a multipole magnetic field generator () comprising four subunits (A,B), each having a core (A,B) with a surrounding coil (A,B) and with a top surface (A,B), wherein all top surfaces of said subunits are preferably arranged in the same plane and wherein all cores are substantially parallel to each other. 1. A method for processing a medium comprising target particles , said method comprising the following steps:a) providing a cartridge having a processing chamber;b) providing a multipole magnetic field generator having at least four subunits, each subunit having a core with a top surface and a coil surrounding said core, wherein the top surfaces of all subunits are arranged adjacent to the cartridge;c) filling the processing chamber with the medium comprising the target particles, wherein the filling is done before, during, and/or after adding magnetic particles (M) to the medium;d) controlling the multipole magnetic field generator such that a time-variable magnetic field is generated that manipulates the magnetic particles and thus mixes the medium;e) binding target particles of the medium to the magnetic particles;f) controlling the multipole magnetic field generator such that the magnetic particles are attracted to a surface of the processing chamber and removing the remaining medium from the processing chamber.2. A processing device for processing a medium comprising target particles , comprising:a cartridge with a processing chamber in which the medium can be provided;a multipole magnetic field generator with at least four subunits, each subunit having a core with a ...

NUCLEIC ACIDS COMPRISING FORMULA (NuGlXmGnNv)a AND DERIVATIVES THEREOF AS IMMUNOSTIMULATING AGENT/ADJUVANT

The present invention relates to nucleic acids of the general formula (I): (NGXGN)and derivatives thereof as an immunostimulating agent/adjuvant and to compositions containing same, optionally comprising an additional adjuvant. The present invention furthermore relates to a pharmaceutical composition or to a vaccine, each containing nucleic acids of formula (I) above and/or derivatives thereof as an immunostimulating agent, and optionally at least one additional pharmaceutically active component, e.g. an antigenic agent. The present invention relates likewise to the use of the pharmaceutical composition or of the vaccine for the treatment of cancer diseases, infectious diseases, allergies and autoimmune diseases etc. Likewise, the present invention includes the use of nucleic acids of the general formula (I): (NGXGN)and/or derivatives thereof for the preparation of a pharmaceutical composition for the treatment of such diseases. 4. Nucleic acid according to claim 3 , characterized in that the nucleic acid molecule according to formula (II) is specifically a nucleic acid molecule according to formula (IIa):{'br': None, 'sub': u', 'l', 'm', 'n', 'v', 'a, 'poly(X)(NGXGN)'}5. Nucleic acid according to claim 3 , characterized in that the nucleic acid molecule according to formula (II) is specifically a nucleic acid molecule according to formula (IIb):{'br': None, 'sub': u', 'l', 'm', 'n', 'v', 'a, 'poly(X)(NGXGN)poly(X)'}6. Nucleic acid according to claim 3 , characterized in that the homopolymeric stretch of nucleic acids poly(X) is a single-stranded claim 3 , a partially double-stranded or a double-stranded RNA sequence.7. Nucleic acid according to claim 1 , characterized in that the homopolymeric stretch of nucleic acids poly(X) is selected from a single-stranded stretch of cytidines (poly(C)) claim 1 , guanosines (poly(G)) claim 1 , adenosines (poly(A)) claim 1 , uridines (poly(U)) claim 1 , inosines (poly(I)) claim 1 , or from a homopolymeric double-stranded stretch ...

Methods for purification of messenger rna

The present invention provides, among other things, methods of purifying messenger RNA (mRNA) including the steps of (a) precipitating mRNA from an impure preparation; (b) subjecting the impure preparation comprising precipitated mRNA to a purification process involving membrane filtration such that the precipitated mRNA is captured by a membrane; and (c) eluting the captured precipitated mRNA from the membrane by re-solubilizing the mRNA, thereby resulting in a purified mRNA solution. In some embodiments, a purification process involving membrane filtration suitable for the present invention is tangential flow filtration.

NOVEL LIPID FORMULATIONS FOR NUCLEIC ACID DELIVERY

The present invention provides novel, stable lipid particles comprising one or more active agents or therapeutic agents, methods of making the lipid particles, and methods of delivering and/or administering the lipid particles. More particularly, the present invention provides stable nucleic acid-lipid particles (SNALP) comprising a nucleic acid (such as one or more interfering RNA), methods of making the SNALP, and methods of delivering and/or administering the SNALP. 1. A nucleic acid-lipid particle comprising:(a) a nucleic acid;(b) a cationic lipid;(c) a non-cationic lipid comprising up to 55 mol % of the total lipid present in the particle, wherein the non-cationic lipid comprises a neutral lipid component comprising a phospholipid of from 3 mol % to 15 mol % of the total lipid present in the particle; and(d) a conjugated lipid that inhibits aggregation of particles comprising from 0.1 mol % to 2 mol % of the total lipid present in the particle.2. The nucleic acid-lipid particle of claim 1 , wherein the nucleic acid comprises an RNA.3. The nucleic acid-lipid particle of claim 2 , wherein the RNA comprises an mRNA.4. The nucleic acid-lipid particle of claim 1 , wherein the phospholipid comprises from 4 mol % to 12 mol % of the total lipid present in the particle.5. The nucleic acid-lipid particle of claim 1 , wherein the phospholipid comprises from 6 mol % to 12 mol % of the total lipid present in the particle.6. The nucleic acid-lipid particle of claim 1 , wherein the neutral lipid component further comprises cholesterol or a derivative thereof.7. The nucleic acid-lipid particle of claim 6 , wherein the cholesterol or derivative thereof comprises from 25 mol % to 45 mol % of the total lipid present in the particle.8. The nucleic acid-lipid particle of claim 6 , wherein the cholesterol or derivative thereof comprises from 40 mol % to 45 mol % of the total lipid present in the particle.9. The nucleic acid-lipid particle of claim 1 , wherein the conjugated lipid ...

NOVEL TETRAZINES AND METHOD OF SYNTHEZISING THE SAME

Provided herein, inter alia, are compositions and methods of synthesis and detection of tetrazines and diazonorcaradienes. 3. The method of claim 2 , wherein said contacting is in a cell.4. The method of claim 3 , further comprising detecting said diazonorcaradiene.5. The method of claim 4 , wherein one of R claim 4 , Rand Ris a detectable moiety.6. The method of claim 5 , wherein said detectable moiety is a fluorophore claim 5 , a radionuclide claim 5 , or a magnetic contrast agent.7. The method of claim 4 , wherein one of R claim 4 , Rand Ris a biomolecule moiety.8. The method of claim 7 , wherein said biomolecule moiety is attached to a detectable moiety.9. The method of claim 8 , wherein said biomolecule moiety is a nucleotide moiety claim 8 , a polynucleotide moiety claim 8 , an amino acid moiety claim 8 , a polypeptide moiety claim 8 , a protein moiety claim 8 , a glycan moiety claim 8 , a lipid moiety claim 8 , a glycolipid moiety or a polysaccharide moiety.10. The method of claim 9 , wherein said biomolecule is RNA moiety or DNA moiety.1112-. (canceled)13. The method of claim 9 , wherein said biomolecule is a protein moiety or polypeptide moiety.1518-. (canceled)19. The method of claim 14 , wherein Ris methyl.20. The method of claim 14 , wherein Ris a detectable moiety.21. The method of claim 20 , wherein said detectable moiety is a fluorophore claim 20 , a radionucleotide claim 20 , or a magnetic contrast agent.22. The method of claim 14 , wherein Ris a biomolecule moiety.23. The method of claim 22 , wherein said biomolecule is a nucleotide moiety claim 22 , a polynucleotide moiety claim 22 , an amino acid moiety claim 22 , a polypeptide moiety claim 22 , a protein moiety claim 22 , a glycan moiety claim 22 , a lipid moiety claim 22 , a glycolipid moiety claim 22 , or a polysaccharide moiety.24. The method of claim 23 , wherein said biomolecule is RNA moiety or DNA moiety.2526-. (canceled)27. The method of claim 23 , wherein said biomolecule is a protein ...

LIGAND CLUSTERS AND METHODS OF THEIR USE AND PREPARATION

Disclosed are compounds of formula (I): Y—X-L-Z, (I) or a salt thereof, where p is 1 to 5; X is a monosaccharide; each Y is independently -L-T, H, protecting group, optionally substituted hydrocarbon, or optionally substituted heteroorganic group, wherein each T is independently a ligand or a protected ligand, and each Lis independently a covalent linker; L2 is a conjugation linker; Z is a therapeutically active agent, protecting group, or a conjugation moiety. Also disclosed are methods of use of the compounds of the invention and methods of their preparation. 2. The compound of claim 1 , or a salt thereof claim 1 , wherein the monosaccharide is a pentose or hexose claim 1 , wherein claim 1 ,when the monosaccharide is a pentose, p is 1 to 3, andwhen the monosaccharide is a hexose, p is 1 to 4.3. The compound of or claim 1 , or a salt thereof claim 1 , wherein the monosaccharide is N-acetylgalactosamine claim 1 , galactosamine claim 1 , galactose claim 1 , mannose claim 1 , allose claim 1 , altrose claim 1 , glucose claim 1 , gulose claim 1 , idose claim 1 , talose claim 1 , arabinose claim 1 , lyxose claim 1 , ribose claim 1 , or xylose.4. The compound of claim 3 , or a salt thereof claim 3 , wherein the monosaccharide is N-acetylgalactosamine.7. The compound of or claim 3 , or a salt thereof claim 3 , wherein each Qis independently absent claim 3 , —CO— claim 3 , —NH— claim 3 , —O— claim 3 , —S— claim 3 , —SO— claim 3 , —CH— claim 3 , —C(O)O— claim 3 , —OC(O)— claim 3 , —C(O)NH— claim 3 , —NHC(O)— claim 3 , —OP(O)(OH)O— claim 3 , or —OP(S)(OH)O—8. The compound of or claim 3 , or a salt thereof claim 3 , wherein each Qis independently NHC(O)— or —C(O)NH—.12. The compound of any one of to claim 3 , or a salt thereof claim 3 , wherein Z is a therapeutically active agent.13. The compound of claim 12 , or a salt thereof claim 12 , wherein the therapeutically active agent is a therapeutically active oligonucleotide.14. The compound of claim 13 , or a salt thereof claim ...

COMPOSITIONS AND METHODS FOR TEMPLATE-FREE GEOMETRIC ENZYMATIC NUCLEIC ACID SYNTHESIS

Disclosed are compositions and methods for template-free nucleic acid synthesis using N-mers and/or anchor primers that comprise at least one XNA or a combination of RNA and DNA. 1. A composition comprising a nucleic acid (NA) sequence capable of participating in geometric synthesis or parallel synthesis reactions , wherein the nucleic acid sequence comprises an arbitrary length , wherein the nucleic acid sequence comprises at least one xenonucleic acid.2. The composition of claim 1 , comprising a plurality of NA sequences claim 1 , wherein each NA sequence of the plurality of NA sequences comprises or consists of a 3-nucleotide oligonucleotide (a 3-mer) claim 1 , a 4-nucleotide oligonucleotide (a 4-mer) claim 1 , or a 5-nucleotide oligonucleotide (a 5-mer) claim 1 , wherein at least one NA sequence in the plurality of NA sequences comprises at least one xenonucleic acid.3. The composition of claim 2 , wherein each NA sequence of the plurality of NA sequences comprises or consists of a 3-mer and wherein the plurality of NA sequences comprises 64 unique NA sequences.4. The composition of claim 2 , wherein each NA sequence of the plurality of NA sequences comprises or consists of a 4-mer and wherein the plurality of NA sequences comprises 256 unique NA sequences.5. The composition of claim 2 , wherein each NA sequence of the plurality of NA sequences comprises or consists of a 5-mer and wherein the plurality of NA sequences comprises 1024 unique NA sequences.6. The composition of claim 2 , wherein the plurality of NA sequences comprises or consists of each unique 3-mer NA sequence claim 2 , each unique 4-mer NA sequence and each unique 5-mer NA sequence claim 2 , and the plurality of NA sequences comprises 1344 unique NA sequences.7. The composition of any one of - claim 2 , wherein each NA sequence of the plurality of NA sequences comprises a monophosphate at a 3′ end or at a 5′ end of the NA sequence.8. The composition of any one of - claim 2 , wherein each NA ...

Human Oncostatin M Antibodies and Methods of Use

Antibodies and compositions capable of neutralizing oncostatin M biological functions are useful in treating diseases and disorders associated with oncostatin M, such as osteoarthritis and idiopathic pulmonary fibrosis. 1. An isolated antibody that specifically binds to human OSM protein and modulates the interaction between human OSM protein and human gp130 protein , comprising a heavy chain complementarity determining region 3 (H-CDR3) selected from the amino acid sequence of SEQ ID NOS: 20 and 21.2. The antibody of claim 1 , further comprising a heavy chain framework 1 sequence claim 1 , H-CDR1 sequence claim 1 , framework 2 sequence claim 1 , H-CDR2 sequence claim 1 , and framework 3 sequence having the amino acid sequence of SEQ ID NO: 48.3. The antibody of claim 2 , wherein the heavy chain residue S31 contacts the OSM protein having the amino acid sequence of SEQ ID NO: 11 at position Q20 and heavy chain residues T30 and Y52 contact the OSM protein having the amino acid sequence of SEQ ID NO: 11 at position G120.4. The antibody of claim 2 , wherein residues of the human variable light chain framework are derived from a human Vkappa germline gene.5. The antibody of claim 4 , wherein the Vkappa germline gene is an IGKV4 germline gene sequence.6. The antibody of claim 2 , further comprising a light chain variable region comprising SEQ ID NO: 8 claim 2 , wherein Xis Y claim 2 , S claim 2 , or A; Xis K claim 2 , E claim 2 , or N; Xis Y claim 2 , W or F; and Xis W.7. The antibody of claim 6 , wherein the L-CDR3 sequence comprises an amino acid sequence of SEQ ID NO: 29.8. The antibody of claim 7 , wherein the L-CDR3 sequence comprises an amino acid sequence selected from the group consisting of SEQ ID NOS: 27 claim 7 , 28 claim 7 , 45 claim 7 , and 46.9. The antibody of claim 6 , wherein the L-CDR1 sequence comprises an amino acid sequence selected from the group consisting of SEQ ID NOS: 23-25 and 30-41.10. The antibody of claim 9 , wherein the L-CDR2 sequence ...

OLIGONUCLEOTIDES AND USE THEREOF

The invention relates to oligonucleotides comprising, or consisting of, the nucleotide sequence SEQ ID NO 4 or to oligonucleotides having nucleotide sequences homologous thereto; or to oligonucleotides having a nucleotide sequence complementary to the nucleotide sequence SEQ ID NO 4 or to oligonucleotides having nucleotide sequences homologous to the complementary nucleotide sequence; or to oligonucleotides having, in view of the reading direction 5′→3′, a nucleotide sequence having an opposite reading direction to the nucleotide sequence SEQ ID NO 4, or to oligonucleotides having nucleotide sequences homologous to the opposite reading direction nucleotide sequence: 112.-. (canceled)20. The oligonucleotide of claim 13 , having a nucleotide sequence of SEQ ID NO 3 or of SEQ ID 4 or of SEQ ID NO 10 or of SEQ ID NO 11 claim 13 , having one or more markings at one or at both of their 5′ ends and/or 3′ ends.21. The oligonucleotide of claim 20 , wherein the one or more markings are markers.22. The oligonucleotide of claim 21 , wherein the one or more markings are fluorescence markers or quenchers.23. The oligonucleotide of claim 13 , having a nucleotide sequence of SEQ ID NO 3 or of SEQ ID 4 or of SEQ ID NO 5 or of SEQ ID NO 10 or of SEQ ID NO 11 or of SEQ ID NO 14 or of SEQ ID NO 15 or of SEQ ID NO 16 in combination with one or more further oligonucleotides selected from forward primers and reverse primers.26. The oligonucleotide of claim 24 , wherein primer (1) is a forward primer and/or primer (2) is a reverse primer; or wherein primer (2) is a forward primer claim 24 , and/or primer (1) is a reverse primer.29. The oligonucleotide of claim 27 , wherein primer (1) is a forward primer and/or primer (2) is a reverse primer; or wherein primer (2) is a forward primer claim 27 , and/or primer (1) is a reverse primer.32Trypanosoma cruziTrypanosoma cruzi. A method of amplifying a pathogen nucleotide sequences claim 13 , preferably for the exclusive amplification of DNA ...