АДЕНОАССОЦИИРОВАННЫЕ ВИРУСНЫЕ ВЕКТОРЫ ДЛЯ ЛЕЧЕНИЯ ЛИЗОСОМНЫХ БОЛЕЗНЕЙ НАКОПЛЕНИЯ

Изобретения относятся к аденоассоциированным вирусным векторам и содержащим их фармацевтическим композициям для лечения лизосомных болезней накопления, и в частности для лечения мукополисахаридозов типа IIIВ (MPSIIIB). Предложен рекомбинантный AAV9-вектор, содержащий промотор CAG, SEQ ID NO: 4, связанный с нуклеотидной последовательностью, кодирующей альфа-N-ацетилглюкозаминидазу, SEQ ID NO: 1, причем нуклеотидная последовательность, кодирующая альфа-N-ацетилглюкозаминидазу, SEQ ID NO: 1, представляет собой последовательность SEQ ID NO: 3, SEQ ID NO: 19 или SEQ ID NO: 22 и последовательность полиА, вставленную между первым концевым повтором AAV и вторым концевым повтором AAV. Предложена также плазмида, содержащая нуклеотидную последовательность, кодирующую альфа-N-ацетилглюкозаминидазу, SEQ ID NO: 1. Причем указанная плазмида представляет собой либо pAAV-CAG-cohNaglu с номером доступа DSM 26626, содержащая нуклеотидную последовательность SEQ ID NO: 3, кодирующую альфа-N-ацетилглюкозаминидазу ...

СПОСОБ ОПРЕДЕЛЕНИЯ МОДУЛИРУЮЩЕГО ДЕЙСТВИЯ ВЕЩЕСТВА НА ЗАВИСИМЫЙ ОТ РЕЦЕПТОРА ИНТЕРЛЕЙКИНА-5 ПУТЬ ПЕРЕДАЧИ СИГНАЛОВ В ЧЕЛОВЕЧЕСКОЙ КЛЕТКЕ ИЛИ КЛЕТКЕ ЖИВОТНОГО

Изобретение относится к биотехнологии и может быть использовано для выявления веществ с фармакологическим действием. Действие вещества на зависимый от рецептора интерлейкина-5 путь передачи сигналов в человеческой клетке или клетке животного определяют путем инкубирования клеток, трансформированных рекомбинантной ДНК, содержащей репортерный ген и регуляторную последовательность, и экспрессирующих функциональный рецептор интерлейкина-5, с исследуемым веществом и последующего определения концентрации продукта репортерного гена. Избирательность действия исследуемого вещества в отношении зависимого от рецептора нитерлейкина-5 пути передачи сигналов определяют путем инкубирования вещества в идентичных условиях с контрольными клетками, экспрессирующими контрольный рецептор, отличный от рецептора интерлейкина-5, и трансформированными рекомбинантной ДНК, содержащей рецепторный ген и регуляторную последовательность, реагирующую на изменение концентрации вторичного переносчика зависимого от контрольного ...

СПОСОБ ПОЛУЧЕНИЯ РЕКОМБИНАНТНЫХ БЕЛКОВ В МОЛОЧНОЙ ЖЕЛЕЗЕ НЕ ТРАНСГЕННЫХ МЛЕКОПИТАЮЩИХ

... 1. Способ получения гетерологичных белков в молоке млекопитающих, кроме человека, опосредованный трансформацией эпителиальных клеток молочной железы (MGE) аденовирусными векторами, который включает следующие этапы: а) индукцию лактации млекопитающего на ранних стадиях его половой зрелости; b) удаление молока и тщательную промывку молочной железы перед инфузией вируса; с) инфузию через канал соска до полного заполнения всей емкости молочной железы раствором, содержащим аденовирусные векторы, несущие гены, кодирующие интересующие гетерологичные белки; d) опорожнение молочной железы в течение 4-24 ч после инфузии. e) сбор молока, содержащегося в молочной железе, начиная через 48 ч после инфузии; f) очистка из молока интересующих гетерологичных белков. 2. Способ по п.1, в котором аденовирусный вектор содержит большинство аденовирусных генов. 3. Способ по п.1, в котором аденовирусный вектор представляет собой аденовирус, лишенный большинства или всех аденовирусных генов, и для образования вирусных ...

ИНСЕРЦИЯ ТАРГЕТНОГО ГЕНА ДЛЯ УЛУЧШЕННОЙ КЛЕТОЧНОЙ ИММУНОТЕРАПИИ

ДВОЙНОЙ ВЕКТОР ДЛЯ ПОДАВЛЕНИЯ ВИРУСА ИММУНОДЕФИЦИТА ЧЕЛОВЕКА

КОНСТРУКЦИИ НУКЛЕИНОВОЙ КИСЛОТЫ И ВЕКТОРЫ ДЛЯ ГЕНОТЕРАПИИ ДЛЯ ПРИМЕНЕНИЯ ДЛЯ ЛЕЧЕНИЯ БОЛЕЗНИ ВИЛЬСОНА И ДРУГИХ СОСТОЯНИЙ

ВИРУСНЫЕ ВЕКТОРЫ, КОДИРУЮЩИЕ РЕКОМБИНАНТНЫЕ ВАРИАНТЫ FIX С ПОВЫШЕННОЙ ЭКСПРЕССИЕЙ, ДЛЯ ГЕНОТЕРАПИИ ГЕМОФИЛИИ В

ГУМАНИЗИРОВАННАЯ МОДЕЛЬ НАРУШЕНИЙ СО СТОРОНЫ ПОЧЕК И ПЕЧЕНИ

ГЕННАЯ ТЕРАПИЯ

СПОСОБЫ ПОЛУЧЕНИЯ ЦЕЛЕВОЙ МОЛЕКУЛЫ В ТРАНСГЕННОМ ЖИВОТНОМ И ВЫДЕЛЕНИЯ ЦЕЛЕВОЙ МОЛЕКУЛЫ

... 1. Трансгенная система для выделения целевого полипептида, обладающего связываемым эпитопом, система, включающая трансгенное животное, имеющее в своем геноме нуклеиновую кислоту, кодирующую поливалентный связывающий полипептид под контролем промотора, который направляет экспрессию в эпителиальных клетках молочной железы, где поливалентный связывающий полипептид включает первую связывающую группу, которая специфически связывает связываемый эпитоп целевого полипептида, и вторую связывающую группу, которая специфически связывает матрикс, посредством чего такой трансгенный поливалентный связывающий полипептид экспрессируется в высоких концентрациях в молоко трансгенного животного, и матрикс, с которым вторая связывающая группа поливалентного связывающего полипептида специфически связывается. 2. Система по п.1, в которой связываемый эпитоп целевого полипептида является удаляемым. 3. Система по п.2, в которой связывающая группа поливалентного связывающего полипептида удаляет связываемый эпитоп ...

МОДЕЛИ РЕТИНОШИЗИСА НА ЖИВОТНЫХ, ОТЛИЧНЫХ ОТ ЧЕЛОВЕКА

IN PROSTATA AKTIVER GEWEBESPEZIFISCHER ENHANCER

REGULIERENDE SEQUENZEN DES VILLIN-GENS AUS DER MAUS UND VERWENDUNG ZUR HERSTELLUNG VON TRANSGENEN TIEREN

AVIÄRES EXPRESSIONSSYSTEM FÜR FREMDPROTEINE

METHODEN ZUR IDENTIFIZIERUNG VON ZUSAMMENSETZUNGEN, DIE FÜR DIE BEHANDLUNG VON FETTLEIBIGKEIT NÜTZLICH SIND, UNTER VERWENDUNG VON FOXC2

TRANSGENE AMPHIBISCHE TIERMODELLE FÜR LYMPHANGIOGENESE

Protein expression

Treatment of retinitis pigmentosa

Gene Therapy

TRANSFORMED CELL LINES

Ex vivo and in vivo expression of the thrombomodulin gene for the treatment of cardiovascular and peripheral vascular diseases

The present invention relates to methods and compositions for treatment of cardiovascular and peripheral vascular diseases using a ex vivo and in vivo gene delivery technologies. One aspect of the present invention relates to a method for treating a vascular disease by introducing a DNA sequence encoding a TM protein or its variant into a segment of a blood vessel ex vitro using a gutless adenovirus vector. Another aspect of the present invention is to provide a method to deliver a gutless adenovirus vector carrying a DNA sequence encoding a TM protein or its variant using a stent.

Delivery system

Regulation of endogenous gene expression in cells using zinc finger proteins

NOVEL ENTITIES FOR CANCER THERAPY

Retroviral delivery sytem

A retroviral delivery system capable of transducing a target site is described. The retroviral delivery system comprises a first nucleotide sequence coding for at least a pan of an envelope protein; and one or more other nucleotide sequences derivable from a retrovirus that ensure transduction of the target site by the retroviral delivery system; wherein the first nucleotide sequence is heterologous with respect to at least one of the other nucleotide sequences; and wherein the first nucleotide sequence codes for at least a part of a rabies G protein or a mutant, variant, derivative or fragment thereof that is capable or recognising the target site.

Conditional mutants of influenza virus M2 protein

Characterisation of gene function using double stranded RNA inhibition

Theraperutic agent screen

Regulation of endogenous gene expression in cells using zinc finger proteins

Ionizing radiation or diathermy-switched gene therapy vectors and their use in antitumour therapy

Adenoviral vectors encoding interferon and their use in gene therapy

The uses of cell-specific splicing in gene therapy

Characterisation of gene function using double stranded rna inhibition

Horizontal cells

Treatment

Novel modified msp-1 nucleic acid sequences and methods for increasing mrna levels and protein expression in cell systems

Novel modified nucleic acid sequences and methods for increasing mrna levels and protein expression in cell systems

Novel modified nucleic acid sequences and methods for increasing mRNA levels and protein expression in cell systems.

The invention provides modified recombinant nucleic acid sequences (preferably DNA) and methods for increasing the mRNA levels and protein expression of proteins which are known to be, or are likely to be, difficult to express in cell culture systems, mammalian eel culture systems, or in transgenic animals. The preferred "difficult" protein candidates for expression using the recombinant techniques of the invention are those proteins derived from heterologous cells preferably those of lower organisms such as parasites, bacteria, and virus having DNA coding sequences comprising high overall AT content or AT rich regions and/or mRNA instability motifs and/or rare codons relative to the recombinant expression system to be used.

Novel modified MSP-1 nucleic acid sequences and methods for increasing mRNA levels and protein expression in cell systems.

The invention provides modified recombinant nucleic acid sequences (prefarably dna)and methods for increasing the mrna levels and protein expression of malarial surface protein msp-1 which is known to be difficult to express in cell culture systems, mammalian cell culture systems, or in trnasgenic animals. The preferred protein candidates for expression using the recombinant techniques of the invention are msp-1 proteins expressed from dna coding sequences comprising reduced overall at content or at rich regions and/or mrna instability motifs and/or rare codons relative to the native msp-1 gene.

DNA sequences used in the production of recombinant human BSSL/CEL in transgenic non-human mammals, and the produced BSSSL/CEL used in infant formulas.

The present invention relates to a DNA molecule containing ...

Novel modified nucleic acid sequences and methods for increasing mrna levels and protein expression in cell systems

NEW DNA SEQUENCES

Novel modified msp-1 nucleic acid sequences and methods for increasing mrna levels and protein expression in cell systems

GENE EXPRESSION SYSTEM

GENE EXPRESSION SYSTEM

Novel modified MSP-1 nucleic acid sequences and methods for increasing mRNA levels and protein expression in cell systems.

Immunotherapeutic compositions for treating and preventing AIDS ARC and HIV infection

Novel modified nucleic acid sequences and methods for increasing mRNA levels and protein expression in cell systems.

NEW DNA SEQUENCES

GENE EXPRESSION SYSTEM

Novel modified msp-1 nucleic acid sequences and methods for increasing mrna levels and protein expression in cell systems

Novel modified nucleic acid sequences and methods for increasing mrna levels and protein expression in cell systems

UROPLAKIN II PROMOTER FROM PIG AND PROCEDURE FOR THE PRODUCTION OF USEFUL PROTEINS USING THE PROMOTER

MODIFIED TERT PROMOTER WITH IMPROVED TUMOR SPECIFICITY AND STRENGTH AS WELL AS THIS CONTAINING REKOMBINANTER VECTOR

ADENOVIRALE VECTORS FOR THE TREATMENT THE HÄMOPHILIE

ADENOVIRALE VECTORS, SPECIFIC TO CELLS, THE ANDROGEN RECEPTOR EXPRIMIEREN, AND METHODS FOR YOUR USE

IMPROVED SYSTEM FOR THE REGULARIZATION OF THE TRANSGENEXPRESSION

ADJUSTING SEQUENCES OF THE VILLIN GENE FROM THE MOUSE AND USE FOR THE PRODUCTION OF TRANSGENEN ANIMALS

GENE REGULARIZATION IN TRANSGENEN ANIMALS USING A VECTOR ON TRANSPOSONBASIS

INDUKTIERBARE TRANSPOSITION WHEN TRANSGENEN ORGANISM WITH TRANSPOSON VECTOR

RETROVIRALES ADMINISTRATION SYSTEM

NOT HUMAN ONES OF ANIMAL MODELS THE TOLERANCE IN RELATION TO HEPATITIS C VIRUS IMMUNOGEN ONE EXHIBITING

BISPEZIFI ANTIBODIES

MORE CELL-SPECIFICALLY EXPRESSIONS /REPLIKATIONSVEKTOR

PREMATURELY AGING MICE TO THE DEMONSTRATION OF THE ROLE OF THE DNA DAMAGE WITH AGING AND INTERFERENCE INTO THE AGECAUSED PATHOLOGY

GEWEBSSPEZIFI EXPRESSION

THERAPEUTIC RETROVIRUS VECTORS FOR GENE THERAPY

PROMOTER TO EXPRESSION OF FOREIGN GENES IN NEURAL CELLS

SELECTIVE-MAKING A REPLICATION VIRALE VECTOREN

HUMAN CELL-SPECIFIC UROTHELIALE REGULATORI SEQUENCES OF THE TRANSKRIPTION OF THE UROPLAKINS, ITS VECTORS AND USE PROCEDURES

PRODUCTION A TRANSGENEN OF A BIRD OF MEANS CYTOPLASMAINJEKTION

CODON OPTIMIZING SYNTHETIC PLASMIDE

VECTOR AND PROCEDURE FOR THE REACHING OF A HIGH ONE EXPRESSION IN CELLS.

PRODUCTION AND BEING USEFUL NON--HUMAN TRANSGENTIERE FOR PRODUCTION HETEROLOGE ANTIBODIES

NUCLEAR FACTORS, WHICH ARE ASSOCIATED WITH OF THE ADJUSTMENT OF THE TRANSKRIPTION.

PRODUCTION OF PEPTIDEN

GENE THERAPY FOR SOLID TUMORS, PAPILLOME AND WARTS

REKOMBINANTE ADENOVIREN VECTOR AND PROCEDURE FOR THE USE

ANIMAL MODEL WITH CANCER, LUNGENEMPHYSEM AND KARDIOMYOPATHIE

HUMAN PROMOTERS OF THE GABA OF B-RECEPTOR 1

TRANSGENE MICE CAPABLE OF THE PRODUCTION OF HETEROLOGER ANTIBODIES

GENTRANSKRIPTION AND IONIZING RADIATION: METHODS AND COMPOSITIONS

REKOMBINANTES ADENO ASSOCIATED VIRUS

GENOMI DNS FOR HUMAN CHOLESTEROL-7 ALPHA HYDROXYLASE AND PROCEDURE FOR ITS USE

USE THAT ADENOVIRALEN E4 REGION FOR the IMPROVEMENT of the GENEXPRESSION

LYSOMALE PROTEINS, MANUFACTURED IN THE MILK OF NIC-HTM-CREPT TRANSGENEN ANIMALS

HUMAN EPIDERMALER GENE PROMOTER

EXPRESSION OF HETEROLIED GENES IN HUMAN BREAST CELLS, INCLUDING HUMAN BRUSTKARZIONOMZELLEN UNDER CONTROL OF REGULATORI SEQUENCES OF WAP OR MMTV

TRANSGENE ANIMALS, THE APP OF ALLELES WITH THE SWEDISH MUTATION ACCOMMODATING

ADENOVIRUS AAV HERMAPHRODITE VIRUS AS SOON AS PROCEDURES FOR USE OF IT

ADENOVIRALE VECTORS CONTAINING a PROSTATE GLAND a SPECIFIC ANTIGEN (PSA) ßRESPONSE ELEMENTß

NEW SUBSTANCES FOR THE CANCER THERAPY

DEVELOPMENT OF A VECTOR FOR THE REACHING OF GENEXPRESSION IN THE EPIDERMIS OF TRANSGENER ANIMALS

KLONIERUNG CHICKEN ANAEMIA VIRUS DNA

ANTAGONIST OF GROWTH HORMONES

HIGH EXPRESSION VECTOR USING ENHANCER ELEMENT

Improved vectors for gene therapy

Lysosomal proteins produced in the milk of transgenic animals

Mir-21 promoter driven targeted cancer therapy

The invention provides a nucleic acid construct comprising a promoter sequence derived from microRNA-21 (miR-21) linked to a nucleic acid sequence encoding an anti-cancer agent, an example of which is a toxin. The constructs of the invention are particularly useful for treating tumors expressing miR-21.

Materials and Methods for the Treatment of Pathological Neovascularization in the Eye

The subject invention provides materials and methods useful in safely and effectively preventing pathological proliferation of blood vessels. The prevention of the over-proliferation of blood vessels according to the subject invention is particularly advantageous for treatment of certain ocular conditions including age-related macular degeneration (AMD), retinopathy of prematurity (ROP) and diabetic retinopathy. In preferred embodiments, the subject invention provides materials and methods for effective treatment of pathological ocular neovascularization using gene therapy. In a specific embodiment the materials and methods of the subject invention can be used to treat AMD.

Therapeutic regimen for treating cancer

The invention provides a method for treating locally advanced resectable esophageal cancer in a human comprising (a) administering to the human a dose of a pharmaceutical composition comprising (i) a pharmaceutically acceptable carrier and (ii) an adenoviral vector comprising a nucleic acid sequence encoding a human TNF-α and operably linked to a promoter, wherein the dose comprises about 4×10 7 to about 4×10 12 particle units (pu) of adenoviral vector, at least once in a therapeutic period comprising up to about 10 weeks, (b) administering a dose of ionizing radiation to the human over the duration of the therapeutic period, and (c) administering a dose of one or more chemotherapeutics to the human over the duration of the therapeutic period, whereby the locally advanced resectable esophageal cancer in the human is treated.

Reagents and methods for modulating cone photoreceptor activity

The present invention provides reagents and methods for modulating cone photoreceptor activity, and devices for assessment of cone photoreceptor activity.

Materials and method for modifying a biochemical component in a plant

A method of modifying the amount of at least one biochemical component in a plant comprising expressing Qua-Quine Starch (QQS) in the plant, the wild-type of which does not express QQS; a transgenic plant, or part thereof, which comprises and expresses QQS as a transgene and in which the amount of at least one biochemical component is modified; a tissue culture of regenerable cells of the transgenic plant; a vector comprising a nucleotide sequence, which encodes the coding sequence of QQS, operably linked to a non-native promoter, which promotes expression of the nucleotide sequence in a plant, which is other than Arabidopsis ; and a method of producing a food or industrial product from a plant.

Regulated expression systems

The application relates to gene constructs for inducible hepato-specific expression of polynucleotides of interest in response to an inducer agent, said constructs comprising (i) an inducible bi-directional operator-promoter with at least one responsive element to said inducer agent flanked by two hepato-specific promoters acting in divergent manner, (ii) a first nucleotide sequence encoding a transactivator which may be activated by said inducer agent operatively coupled to the first hepato-specific promoter and (iii) a second nucleotide sequence operatively coupled to the second hepato-specific promoter, wherein the promoters are induced as a consequence of the binding of the transactivator to the operator region of the operator-promoter in the presence of the inducer agent.

Method and Composition to Increase Radiation-Induced Tumor Therapeutic Effects

Disclosed herein are methods and compositions for treating cancer by increasing radiation-induced damage to cancer without increasing radiation-induced side effects by increasing secretory ASMase levels specifically in tumor endothelium, and inducing apoptosis of tumor endothelial cells by treating the tumor with radiation. ASMase levels are increased in tumor endothelium by administration of a recombinant DNA construct comprising a region coding for a functional ASMase linked to particular transcriptional regulatory sequences that confer tissue-specific expression of ASMase.

Novel Therapeutical Tools and Methods for Treating Blindness

The present inventions relates to the use of an isolated nucleic acid molecule comprising a nucleotide sequence coding for a hyperpolarizing light-gated ion channel or pump gene from an archeon or for a light-active fragment of said gene, or the nucleotide sequence complementary to said nucleotide sequence, for treating or ameliorating blindness. The light-gated ion channel or pump gene can be a halorhodopsin gene.

Tumor-specific promoter and oncolytic virus vector comprising the same

Provided are tumor-specific promoters, oncolytic virus vectors and a pharmaceutical composition comprising the virus vector. The virus vector comprising the novel tumor-specific promoter shows excellent oncolytic effects on tumor cells, and thus it is useful for treating a cancer.

Recombinant tumor vaccine and method of producing such vaccine

The present disclosure provides tumor vaccines useful for preventing and treating tumors and cancers. The tumor vaccines may contain nucleic acids encoding for antigen presenting peptides, cytokines and other factors useful for preventing and treating tumors and cancers, or expression vectors or viruses containing such nucleic acids, or host cells containing such nucleic acids or expression vectors.

Nucleic acid construct systems capable of diagnosing or treating a cell state

Nucleic acid construct systems are disclosed capable of diagnosing and treating a cell state (e.g. disease state). Methods of diagnosing and treating disease states using the nucleic acid constructs described herein are also disclosed. In addition, methods of screening for agents capable of reversing a disease phenotype using the nucleic acid constructs of the present invention are disclosed.

Production of Transgenic Avians Using Improved Retroviral Vectors

A transgenic avian containing in its genome an exogenous nucleotide sequence which includes a promoter component and a vector with reduced promoter interference wherein the exogenous nucleotide sequence is integrated into the genome and the avian.

Method for expression of small antiviral rna molecules with reduced cytotoxicity within a cell

In one aspect, the invention provides methods and compositions for the expression of small RNA molecules within a cell using a retroviral vector (FIG. 1 A). Small interfering RNA (siRNA) can be expressed using the methods of the invention within a cell. In a further aspect, the invention provides methods for producing siRNA encoding lentivirus where the siRNA activity may interfere with the lentiviral life cycle. In yet a further aspect, the invention provides methods for expression of a small RNA molecule within a cell, such as an siRNA capable of downregulating CCR5, wherein expression of the small RNA molecule is relatively non-cytotoxic to the cell. The invention also includes small RNA molecules, such as an siRNA capable of downregulating CCR5, that are relatively non-cytotoxic to cells.

Chimeric gene constructs for generation of fluorescent transgenic ornamental fish

Four zebrafish gene promoters, which are skin specific, muscle specific, skeletal muscle specific and ubiquitously expressed respectively, were isolated and ligated to the 5′ end of the EGFP gene. When the resulting chimeric gene constructs were introduced into zebrafish, the transgenic zebrafish emit green fluorescence under a blue light or ultraviolet light according to the specificity of the promoters used. Thus, new varieties of ornamental fish of different fluorescence patterns, e.g., skin fluorescence, muscle fluorescence, skeletal muscle-specific and/or ubiquitous fluorescence, are developed.

Expression vectors comprising engineered genes

The invention provides expression vectors, nucleic acids, vector particles and methods of treatment involving these vector particles, comprising an engineered KCNA1 gene encoding an edited Kv1.1 potassium channel, as well as methods of confirming the presence of engineered KCNA1 mRNA in a cell. The features of the engineered KCNA1 gene combine to advantageously enhance the translation and activity of the Kv1.1 protein and improve detection of KCNA1 gene expression in a cell and can be used for example in the treatment of epilepsy and similar neurological disorders.

Optimised Coding Sequence and Promoter

An optimized coding sequence of human blood clotting factor eight (VIII) and a promoter may be used in vectors, such as rAAV, for introduction of factor VIII, and/or other blood clotting factors and transgenes. Exemplary of these factors and transgenes are alpha-1-antitrypsin, as well as those involved in the coagulation cascade, hepatocyte biology, lysosomal storage, urea cycle disorders, and lipid storage diseases. Cells, vectors, proteins, and glycoproteins produced by cells transformed by the vectors and sequence, may be used in treatment. 1. An isolated nucleic acid molecule comprising a nucleotide sequence having at least 75% homology to the nucleotide sequence of SEQ ID NO: 1 and which encodes functional factor FVIII.2. The nucleic acid molecule of having at least 85% homology to the nucleotide sequence of SEQ ID NO: 1.3. The nucleic acid molecule of claim 1 , which encodes for a protein comprising the sequence of SEQ ID NO: 2 or SEQ ID NO: 21 having between 0 and 10 amino acid changes thereto.4. The nucleic acid molecule of claim 3 , which encodes for a protein comprising the sequence of SEQ ID NO: 2 or SEQ ID NO: 21.5. The nucleic acid molecule of comprising a nucleotide sequence selected from the sequence of SEQ ID NO: 1 claim 1 , SEQ ID NO: 4 claim 1 , SEQ ID NO: 5 claim 1 , SEQ ID NO: 6 and SEQ ID NO: 7.6. The nucleic acid molecule of further comprising a nucleotide sequence having at least 85% homology to the nucleotide sequence of SEQ ID NO: 3.7. A vector comprising a nucleic acid molecule of .8. A host cell comprising the nucleic acid molecule of or the vector of .9. A protein or glycoprotein expressed by the host cell of .10. (canceled)11. A method of treating haemophilia comprising administering a vector according to to a patient suffering from haemophilia.12. (canceled)13. (canceled)14. A method for delivery of a nucleotide sequence encoding a functional factor VIII to a subject claim 1 , which method comprises administering to the said subject a ...

METHODS OF USE OF INHIBITORS OF PHOSPHODIESTERASES AND MODULATORS OF NITRIC OXIDE, REACTIVE OXYGEN SPECIES, AND METALLOPROTEINASES IN THE TREATMENT OF PEYRONIE'S DISEASE, ARTERIOSCLEROSIS AND OTHER FIBROTIC DISEASES

The present methods and compositions are of use for treatment of conditions involving fibrosis, such as Peyronie's disease plaque, penile corporal fibrosis, penile veno-occlusive dysfunction, Dupuytren's disease nodules, vaginal fibrosis, clitoral fibrosis, female sexual arousal disorder, abnormal wound healing, keloid formation, general fibrosis of the kidney, bladder, prostate, skin, liver, lung, heart, intestines or any other localized or generalized fibrotic condition, vascular fibrosis, arterial intima hyperplasia, atherosclerosis, arteriosclerosis, restenosis, cardiac hypertrophy, hypertension or any condition characterized by excessive fibroblast or smooth muscle cell proliferation or deposition of collagen and extracellular matrix in the blood vessels and/or heart. In certain embodiments, the compositions may comprise a PDE-4 inhibitor, a PDE-5 inhibitor, a compound that elevates cGMP and/or PKG, a stimulator of guanylyl cyclase and/or PKG, a combination of a compound that elevates cGMP, PKG or NO with an antioxidant that decreases ROS, or a compound that increases MMP activity. 1. A method comprising administering a cyclic guanosine 3′ , 5′-monophosphate (cGMP) type 5 phosphodiesterase (PDE-5) inhibitor according to a continuous , long-term regimen to an individual with at least one of a penile tunical fibrosis and corporal tissue fibrosis , wherein the PDE-5 inhibitor is selected from the group consisting of sildenafil , tadalafil , and vardenafil , wherein the PDE-5 inhibitor is administered at a dosage of up to 1.5 mg/kg/day , and wherein the PDE′″5 inhibitor is administered for greater than or equal to 45 days.2. The method of claim 1 , wherein the PDE-5 inhibitor is sildenafil.3. The method of claim 1 , wherein the PDE-5 inhibitor is tadalafil.4. The method of claim 1 , wherein the PDE-5 inhibitor is vardenafil.5. The method of claim 1 , wherein the individual has penile tunical fibrosis.6. The method of claim 1 , wherein the individual has corporal ...

Genetically modified mesenchymal stem cells expressing alpha-1 antitrypsin (aat)

Genetically modified mesenchymal stem cells can be used as a medicament in the treatment of medical conditions associated with inflammation and/or an unwanted immune response in subjects without an alpha1-antitrypsin (AAT) deficiency. The stem cells include an exogenous nucleic acid, which includes (i) an Alpha-1 antitrypsin (AAT) encoding region operably linked to (ii) a promoter or promoter/enhancer combination.

Cell lines expressing inserted secretable reporter genes at multiple stages of differentiation

A composition of matter comprises one or more cell lines configured to inducibly differentiate to at least a first stage of differentiation and a second, subsequent stage of differentiation. Each of the one or more cell lines are genetically edited to express one or more first stage inserted secretable reporter genes placed under control of promoters for genes canonically expressed during the first stage of differentiation. The cell lines are further genetically edited to express one or more second stage inserted secretable reporter genes placed under control of promoters for genes canonically expressed during the second stage of differentiation, but not during the first stage of differentiation, wherein the one or more second stage inserted secretable reporter genes are different than the one or more first stage inserted secretable reporter genes.

METHODS AND COMPOSITIONS FOR TREATING GENETICALLY LINKED DISEASES OF THE EYE

Expression vectors and therapeutic methods of using such vectors in the treatment of diseases of the eye resulting from failure to produce a specific protein in the eye, or the production of a non-functional protein in the eye. 169-. (canceled)70. An expression vector comprising an expression cassette , wherein the expression cassette comprises a promoter operably-linked to a nucleic acid sequence encoding a therapeutic protein , wherein the level of expression of the therapeutic protein is high enough that the amount of a delivery vehicle comprising the expression vector needed to alleviate symptoms of an eye disease , elicits a minimal immune response when administered to the eye.71. The expression vector of claim 70 , wherein the promoter is an eye-specific promoter.72. The expression vector of claim 70 , wherein the promoter comprises at least a portion of a promoter selected from the group consisting of a retinoschisin promoter claim 70 , a rhodopsin promoter claim 70 , a rhodopsin kinase promoter claim 70 , a CRX promoter claim 70 , and an interphotoreceptor retinoid binding protein (IRBP) promoter73. The expression vector of claim 70 , wherein the expression cassette comprises an enhancer sequence that enhances the activity of the promoter.74. The expression vector of claim 73 , wherein the enhancer sequence is an interphotoreceptor retinoid binding protein enhancer sequence.75. The expression vector of claim 70 , wherein the expression cassette comprises at least a portion of intron 1 of a retinoschisin gene.76. The expression vector of claim 75 , wherein the at least a portion is located within the nucleic acid sequence encoding a therapeutic protein.77. The expression vector of claim 75 , wherein the at least a portion of intron 1 comprises the splice donor and splice acceptor sequences of intron 1 of a retinoschisin gene.78. The expression vector of claim 70 , wherein the therapeutic protein is selected from the group consisting of a retinoschisin protein ...

Synthetic Promoters

CHO cell-specific synthetic promoter constructs for expressing recombinant proteins, a library of promoter constructs thereof, and a method for producing the promoter constructs. The promoter constructs enable precise control of recombinant gene transcription over three orders of magnitude, with the top expressing promoters capable of double the transcriptional activity of the CMV promoter. 1. A CHO cell , comprising a synthetic promoter suitable for eliciting recombinant protein expression therein , said synthetic promoter comprising a promoter core and upstream thereof two or more transcription factor regulatory elements independently selected from the group consisting of NFκB-RE , E-box , AP1 , CRE , GC-Box , E41F , C/EBPα-RE , OCT and RARE.2. The CHO cell according to claim 1 , wherein the promoter core is selected from CMV claim 1 , SV40 claim 1 , UbC claim 1 , EF1A claim 1 , PGK and CAGG.3. The CHO cell according to claim 1 , wherein the synthetic promoter comprises 2 to 50 transcription factor regulatory elements.4. The CHO cell according to claim 1 , wherein the transcription factor regulatory elements are all the same type.5. The CHO cell according to claim 1 , wherein the transcription factor regulatory elements are a combination of different types claim 1 , which are claim 1 , optionally claim 1 , independently selected from NFκB-RE claim 1 , E-box claim 1 , GC-Box claim 1 , C/EBPα-RE claim 1 , CRE and E41F.6. The CHO cell according to claim 1 , wherein the transcription factor regulatory elements are arranged in tandem.7. (canceled)8. The CHO cell according to claim 1 , whereina) synthetic promoter DNA sequence is 0.9 or less of the size of the full length CMV promoter sequence, and/orb) the synthetic promoter has a transcriptional activity per unit DNA sequence thereof which in greater than the transcriptional activity per unit DNA of CMV promoter.9. (canceled)10. The CHO cell according to claim 1 , wherein the CHO cell is selected from CHO-S claim 1 , ...

METHOD FOR EXPRESSION OF SMALL ANTIVIRAL RNA MOLECULES WITH REDUCED CYTOTOXICITY WITHIN A CELL

In one aspect, the invention provides methods and compositions for the expression of small RNA molecules within a cell using a retroviral vector (FIG. A). Small interfering RNA (siRNA) can be expressed using the methods of the invention within a cell. In a further aspect, the invention provides methods for producing siRNA encoding lentivirus where the siRNA activity may interfere with the lentiviral life cycle. In yet a further aspect, the invention provides methods for expression of a small RNA molecule within a cell, such as an siRNA capable of downregulating CCR5, wherein expression of the small RNA molecule is relatively non-cytotoxic to the cell. The invention also includes small RNA molecules, such as an siRNA capable of downregulating CCR5, that are relatively non-cytotoxic to cells. 120.-. (canceled)21. A retroviral construct for expressing an RNA within a cell , comprising:a nucleic acid having the R and U5 sequences from a 5′ lentiviral long terminal repeat (LTR);a self-inactivating lentiviral 3′ LTR; anda first promoter configured to be operably linked to a first RNA coding region encoding a first RNA,wherein the first promoter is located between the 5′ LTR and the 3′ LTR, and wherein the first RNA comprises a RNA duplex.22. The retroviral construct of claim 21 , wherein the first RNA comprises a sequence that is at least about 90% complementary to a target region of a pathogenic virus genome or genome transcript.23. The retroviral construct of claim 22 , wherein the pathogenic virus is human immunodeficiency virus (HIV) claim 22 , hepatitis A virus (HAV) claim 22 , hepatitis B virus (HBV) claim 22 , hepatitis C virus (HCV) claim 22 , cytomegalovirus (CMV) claim 22 , herpes simplex virus (HSV) claim 22 , influenza virus claim 22 , adeno virus claim 22 , human papillomavirus claim 22 , Coxsackieviruses claim 22 , Measles virus claim 22 , or poliovirus.24. The retroviral construct of claim 23 , wherein the pathogenic virus is HIV-1 or HIV-2.25. The ...

METHODS AND COMPOSITIONS FOR PRODUCTION OF AROMATIC AND OTHER COMPOUNDS IN YEAST

The present disclosure includes methods and components for production of valuable industrial compounds in yeast. In an embodiment, the present invention provides a nucleic acid construct with increased stability for gene expression or gene editing comprising: a nucleic acid sequence encoding one or more of SEQ ID NO: 1-8 (CENs 1-8); and one or more regulatory elements functional in a yeast cell. In an embodiment of the present invention the nucleic acid constructs are vectors, preferably episomal vectors. High expression promoters, as well as methods for increasing production of compounds such as aromatics are disclosed. 1. A nucleic acid construct for gene expression or gene editing in yeast comprising:a nucleic acid sequence encoding one or more of SEQ ID NO: 1-8 (CENs 1-8); andone or more regulatory elements functional in a yeast cell.2. The nucleic acid construct of claim 1 , wherein said nucleic acid construct further comprises an autonomously replicating sequence (ARS).3. The nucleic acid construct of claim 1 , wherein said nucleic acid construct includes a multiple cloning site for insertion of gene of interest operably linked to the regulatory elements.4. The nucleic acid construct of claim 1 , wherein the regulatory element is a promoter.5. The nucleic acid construct of claim 1 , wherein said regulatory element is a terminator sequence.6. The nucleic acid construct of wherein the gene of interest is inserted in a multiple cloning site.7. A vector comprising the nucleic acid construct of .8. The vector of claim 7 , wherein said vector is a plasmid vector.9. The vector of claim 7 , wherein said vector is an episomal vector.10. The vector of claim 8 , wherein said plasmid vector has increased stability compared to the same plasmid vector lacking one or more of SEQ ID NO:1-8 (CENs 1-8).11. A cell claim 1 , tissue claim 1 , or organ comprising the nucleic acid construct of .12. The nucleic acid construct of claim 4 , wherein the promoters are selected from the ...

PROMOTERS, EXPRESSION CASSETTES, VECTORS, KITS, AND METHODS FOR THE TREATMENT OF ACHROMATOPSIA AND OTHER DISEASES

The present invention provides isolated promoters, transgene expression cassettes, vectors, kits, and methods for treatment of genetic diseases that affect the cone cells of the retina. 1. An isolated promoter comprising approximately 1.8 kb of the 5′-NTR of the cyclic nucleotide-gated ion channel beta 3 (CNGB3) gene.2. The promoter of comprising the sequence SEQ ID NO: 13. An isolated promoter comprising approximately 1.6 kb of the 5′-NTR of the CNGB3 gene.4. The promoter of comprising SEQ ID NO:2.5. An isolated promoter comprisingapproximately 400 bp of the cytomegalovirus (CMV) enhancer andapproximately 1.4 kb of the 5′-NTR of the CNGB3 gene.6. The promoter of comprising the following sequences:cytomegalovirus (CMV) enhancer set forth as SEQ ID NO: 3 and the 5′-NTR of the CNGB3 gene set forth as SEQ ID NO: 4.7. The promoter of claim 1 , wherein the CNGB3 gene is the human CNGB3 gene.8. The promoter of claim 1 , wherein said promoter is capable of promoting CNGB3 expression in S-cone cells claim 1 , M-cone cells claim 1 , and L-cone cells.9. The promoter of claim 1 , wherein said promoter is capable of promoting cyclic nucleotide-gated ion channel alpha 3 (CNGA3) expression in S-cone cells claim 1 , M-cone cells claim 1 , and L-cone cells.10. The promoter of claim 1 , wherein said promoter is capable of promoting guanine nucleotide binding protein subunit alpha transducin 2 (GNAT2) expression in S-cone cells claim 1 , M-cone cells claim 1 , and L-cone cells.11. A transgene expression cassette comprising{'claim-ref': {'@idref': 'CLM-00001', 'claim 1'}, '(a) the promoter of ;'}(b) a nucleic acid selected from the group consisting of a CNGB3 nucleic acid, a CNGA3 nucleic acid, and a GNAT2 nucleic acid; and(c) minimal regulatory elements.12. A transgene expression cassette comprising{'claim-ref': {'@idref': 'CLM-00001', 'claim 1'}, '(a) the promoter of ,'}(b) a CNGB3 nucleic acid, and(c) minimal regulatory elements.13. (canceled)14. A nucleic acid vector comprising ...

GENE THERAPY FOR TREATING FAMILIAL HYPERCHOLESTEROLEMIA

Compositions and regimens useful in reducing one or more of: LDL-cholesterol, total cholesterol, and/or fasting triglycerides in a subject, and/or modifying fractional catabolic rate (FCR) of LDL apolipoprotein B (apoB) from baseline to a selected time point after rAAV administration are provided. The method involves administering to the human subject via a peripheral vein by infusion of a suspension of replication deficient recombinant adeno-associated virus (rAAV). 1. A pharmaceutical composition suitable for peripheral vein infusion in human subjects , comprising a suspension of replication deficient recombinant adeno-associated virus (rAAV) in a formulation buffer , wherein:(a) the rAAV comprises a vector genome comprising AAV ITRs and a nucleic acid sequence encodes a human LDL receptor (hLDLR) operably linked to a liver specific promoter, said vector genome packaged in an AAV8 capsid;(b) the formulation buffer comprises an aqueous solution of phosphate buffered saline and a Poloxamer; and [{'sup': '13', '(i) the rAAV Genome Copy (GC) titer is at least 1×10GC/ml;'}, '(ii) the rAAV is at least about 95% free of empty capsids as determined by oqPCR or ddPCR;', '(iii) the Empty:Full particle ratio is between 0:4 to 1:4; and', {'sup': '11', '(iv) a dose of 5×10GC/kg of the rAAV suspension decreases baseline cholesterol levels in a double knockout (DKO) LDLR−/− Apobec−/− mouse model of Homozygous Familial Hypercholesterolemia (HoFH) by 25% to 75%.'}], '(c) one or more of2. The composition according to claim 1 , wherein the rAAV is AAV8.TBG.hLDLR.3. The composition according to claim 1 , wherein the formulation buffer is 180 mM NaCl claim 1 , 10 mM Na phosphate claim 1 , 0.001% Poloxamer 188 claim 1 , pH 7.3.4. The composition according to suitable for use in treating a human subject diagnosed with Familial Hypercholesterolemia (FH).5. The composition according to claim 1 , wherein the composition is administrable to the human subject via a peripheral vein by ...

AAV VIRIONS WITH DECREASED IMMUNOREACTIVITY AND USES THEREFOR

Methods of making and using recombinant AAV virions with decreased immunoreactivity are described. The recombinant AAV virions include mutated capsid proteins or are derived from non-primate mammalian AAV serotypes and isolates that display decreased immunoreactivity relative to AAV-2. 143-. (canceled)44. A mutated adeno-associated virus (AAV) capsid protein comprising a substitution of one or more of the amino acids occurring at a position corresponding to a position of the full-length AAV-2 VP2 capsid , wherein the one or more amino acids are selected from the group consisting of amino acid:124 substituted with a threonine;126 substituted with an alanine;127 substituted with an alanine or a leucine;128 substituted with an alanine or an aspartic acid;130 substituted with a threonine or an alanine;131 substituted with a glutamine or an alanine;132 substituted with a glutamic acid, an alanine, or an asparagine;133 substituted with an alanine;134 substituted with a glutamine, a phenylalanine, or an alanine;188 substituted with an alanine;190 substituted with an alanine;191 substituted with a serine;193 substituted with an alanine;245 substituted with an alanine;246 substituted with an alanine;247 substituted with an alanine;248 substituted with an alanine;315 substituted with an alanine;317 substituted with an alanine;318 substituted with an alanine;320 substituted with an alanine;322 substituted with an alanine;329 substituted with an arginine;331 substituted with an alanine;332 substituted with an alanine;334 substituted with an alanine;335 substituted with an alanine;347 substituted with a cysteine;350 substituted with a lysine;354 substituted with an alanine;355 substituted with a threonine or an alanine;356 substituted with an arginine;357 substituted with a glutamic acid, a glutamine, an alanine, or an asparagine;358 substituted with a lysine;359 substituted with an alanine;360 substituted with a histidine, a lysine, or an alanine;361 substituted with an alanine ...

RECOMBINANT CONSTRUCTS AND TRANSGENIC FLUORESCENT ORNAMENTAL FISH THEREFROM

The present invention relates to the method and use of reef coral fluorescent proteins in making transgenic red, green and yellow fluorescent zebrafish. Preferably, such fluorescent zebrafish are fertile and used to establish a population of transgenic zebrafish and to provide to the ornamental fish industry for the purpose of marketing. Thus, new varieties of ornamental fish of different fluorescence colors from a novel source are developed. 171.-. (canceled)73. The transgenic fish of claim 72 , wherein the first and second fluorescent proteins are ZsGreen1 claim 72 , ZsYellow1 claim 72 , DsRed2 claim 72 , GFP claim 72 , eGFP claim 72 , YFP claim 72 , eYFP claim 72 , BFP claim 72 , eBFP claim 72 , CFP claim 72 , eCFP claim 72 , FP claim 72 , AmCyan1 claim 72 , DsRed-Express claim 72 , AsRed2 claim 72 , HcRed1 claim 72 , mPlum claim 72 , mCherry claim 72 , tdTomato claim 72 , mStrawberry claim 72 , J-Red claim 72 , DsRed-monomer claim 72 , mOrange claim 72 , mKO claim 72 , MCitrine claim 72 , Venus claim 72 , Ypet claim 72 , EYFP claim 72 , Emerald claim 72 , CyPet claim 72 , mCFPm claim 72 , Cerulean claim 72 , or T-Sapphire.74. The transgenic fish of claim 73 , wherein the first and second fluorescent proteins are ZsGreen1.75. The transgenic fish of claim 73 , wherein the first and second fluorescent proteins are DsRed 2.76. The transgenic fish of claim 72 , wherein said fish β-actin promoter is a carp β-actin promoter.77. The transgenic fish of claim 72 , wherein said fish myosin light chain promoter is a zebrafish fast skeletal myosin light chain promoter.78. The transgenic fluorescent fish of claim 72 , wherein each of said genes comprise at least two polyadenylation signals positioned in tandem.79. The transgenic fluorescent fish of claim 78 , wherein said polyadenylation signals are viral polyadenylation signals.80. The transgenic fluorescent fish of claim 79 , wherein said viral polyadenylation signals are SV40 polyadenylation sequences.81. The transgenic ...

METHOD AND COMPOSITION TO INCREASE RADIATION-INDUCED TUMOR THERAPEUTIC EFFECTS

Disclosed herein are methods and compositions for treating cancer by increasing radiation-induced damage to cancer without increasing radiation-induced side effects by increasing secretory ASMase levels specifically in tumor endothelium, and inducing apoptosis of tumor endothelial cells by treating the tumor with radiation. ASMase levels are increased in tumor endothelium by administration of a recombinant DNA construct comprising a region coding for a functional ASMase linked to particular transcriptional regulatory sequences that confer tissue-specific expression of ASMase. 1. A method to treat cancer by increasing radiation-induced damage to a tumor without increasing radiation-induced side effects comprising:(1) increasing secretory ASMase levels specifically in tumor endothelium; and(2) inducing apoptosis of tumor endothelial cells by treating the tumor with radiation.2. The method of claim 1 , where the cancer is a solid tumor.3. The method of claim 1 , where the increase in radiation-induced damage to cancer without an increase in radiation-induced side effects is achieved by sensitizing the tumor to radiation.4. The method of claim 1 , where the increase in radiation-induced damage to cancer without an increase in radiation-induced side effects is achieved by sensitizing the angiogenic epithelium of the tumor to radiation.5. The method of claim 1 , wherein secretory ASMase levels are increased specifically in tumor endothelium through the administration of a gene therapy construct.6. The method of claim 5 , wherein the gene therapy construct comprises a recombinant DNA construct comprising a region coding for a functional secretory ASMase linked to transcriptional regulatory sequences that confer tissue-specific expression of the secretory ASMase.7. The method of claim 5 , wherein the gene therapy construct comprises a replication defective adenovirus expression vector comprising a recombinant DNA construct comprising a region coding for a functional ...

METHODS AND COMPOSITIONS FOR GENE DELIVERY TO ON BIPOLAR CELLS

Disclosed are capsid-modified rAAV expression vectors, as well as infectious virions, compositions, and pharmaceutical formulations that include them. Also disclosed are methods of preparing and using novel capsid-protein-mutated rAAV vector constructs in a variety of diagnostic and therapeutic applications including, inter alia, as delivery agents for diagnosis, treatment, or amelioration of one or more diseases, disorders, or dysfunctions of the mammalian eye. Also disclosed are methods for intravitreal delivery of therapeutic gene constructs to retinal neuron cells, and specifically to ON bipolar cells, of the mammalian eye, as well as use of the disclosed compositions in the manufacture of medicaments for a variety of in vitro and/or in vivo applications including the treatment of retinitis pigmentosa, melanoma-associated retinopathy, and congenital stationary night blindness. 1. An adeno-associated viral (AAV) particle comprising:(a) a recombinant adeno-associated viral (rAAV) vector polynucleotide that comprises a nucleic acid segment that encodes a diagnostic or therapeutic agent operably linked to an ON bipolar cell-specific promoter that is capable of expressing the nucleic acid segment in one or more middle retinal neuron cells of a mammalian eye; and(b) a modified capsid protein, wherein the modified capsid protein comprises at least a first non-native amino acid at a position that corresponds to a surface-exposed amino acid residue in the wild-type AAV2 capsid protein, and further wherein the transduction efficiency of a virion comprising the modified capsid protein is higher than that of a virion comprising a corresponding, unmodified wild-type capsid protein.2. The AAV particle of claim 1 , wherein the modified capsid protein comprises three or more non-native amino acid substitutions at positions corresponding to three distinct surface-exposed amino acid residues of the wild-type AAV2 capsid protein as set forth in SEQ ID NO:2; or to three distinct ...

Optimised Coding Sequence and Promoter

An optimized coding sequence of human blood clotting factor eight (VIII) and a promoter may be used in vectors, such as rAAV, for introduction of factor VIII, and/or other blood clotting factors and transgenes. Exemplary of these factors and transgenes arc alpha-1-antitrypsin, as well as those involved in the coagulation cascade, hepatocye biology, lysosomal storage, urea cycle disorders, and lipid storage diseases. Cells, vectors, proteins, and glycoproteins produced by cells transformed by the vectors and sequence, may be used in treatment. 122-. (canceled)23. A recombinant adeno-associated virus (AAV) particle comprising a heterologous nucleic acid sequence and a promoter that is operably linked to and drives expression of said heterologous nucleic acid sequence , wherein said promoter has at least 90% sequence identity to the nucleotide sequence of SEQ ID NO:3.241. The recombinant AAV particle of claim , wherein said promoter is less than 350 base pairs in length.251. The recombinant AAV particle of claim , wherein said promoter comprises the nucleotide sequence of SEQ ID NO:3.261. The recombinant AAV particle of claim , where said promoter consists essentially of the nucleotide sequence of SEQ ID NO:3.271. The recombinant AAV particle of claim , where said promoter consists of the nucleotide sequence of SEQ ID NO:3.281. The recombinant AAV particle of claim which is of AAVS serotype.291. A composition of matter comprising the recombinant AAV particle of claim and a pharmaceutically acceptable carrier. This application is a continuation of U.S. patent application Ser. No. 14/680,836, filed Apr. 7, 2015, which is a divisional of U.S. patent application Ser. No. 13/382,953, filed Apr. 18, 2012, which is a U.S. national phase filing under 35 U.S.C. §371 of International Patent Application No. PCT/US10/41378, filed Jul. 8, 2010, which claims priority to U.K. Application No. 0911870.4 filed Jul. 8, 2009, all of which are incorporated by reference herein in their ...

METHOD OF DETECTING AND/OR IDENTIFYING ADENO-ASSOCIATED VIRUS (AAV) SEQUENCES AND ISOLATING NOVEL SEQUENCES IDENTIFIED THEREBY

Adeno-associated virus rh.20 sequences, vectors containing same, and methods of use are provided. 1. A cultured host cell containing a recombinant nucleic acid molecule encoding an AAV vp1 capsid protein having a sequence comprising amino acids 1 to 738 of SEQ ID NO: 85 (AAVrh.20) , wherein the recombinant nucleic acid molecule further comprises a heterologous non-AAV sequence.2. The cultured host cell according to claim 1 , which further comprises a functional rep gene.3. A cultured host cell containing a recombinant nucleic acid molecule encoding an AAV vp2 capsid protein having a sequence comprising amino acids 138 to 738 of SEQ ID NO: 85 (AAVrh.20) claim 1 , wherein the recombinant nucleic acid molecule further comprises a heterologous non-AAV sequence.4. The cultured host cell according to claim 3 , which further comprises a functional rep gene.5. A cultured host cell containing a recombinant nucleic acid molecule encoding an AAV vp3 capsid protein having a sequence comprising amino acids 204 to 738 of SEQ ID NO: 85 (AAVrh.20) claim 3 , wherein the recombinant nucleic acid molecule further comprises a heterologous non-AAV sequence.6. The cultured host cell according to claim 5 , which further comprises a functional rep gene.7. A cultured host cell containing a recombinant nucleic acid molecule comprising (a) nucleotides 844 to 3057 of SEQ ID NO: 27 claim 5 , (b) nucleotides 1255 to 3057 of SEQ ID NO: 27 claim 5 , or (c) nucleotides 1453 to 3057 of SEQ ID NO: 27 claim 5 , wherein the recombinant nucleic acid molecule further comprises a heterologous non-AAV sequence.8. The cultured host cell according to claim 7 , which further comprises a rep gene.9. The cultured host cell according to claim 2 , wherein the rep gene is from AAV2.10. The cultured host cell according to claim 4 , wherein the rep gene is from AAV2.11. The cultured host cell according to claim 6 , wherein the rep gene is from AAV2.12. The cultured host cell according to claim 8 , wherein the rep ...

Synthetic promoters for high throughput screening and gene modulation

The present invention provides nucleic acid constructs, expression vectors, transgenic cell and methods of making and using the same, wherein the nucleic acid construct includes a synthetic promoter designed from the endogenous promoter of BIRC5 and LAMC2. In illustrative working embodiments of the invention, an exogenous nucleic acid fragment encoding thymidine kinase is operably linked to the synthetic promoter which is then shown to regulate the expression of this polypeptide.

SYNPIII, A PROMOTER FOR THE SPECIFIC EXPRESSION OF GENES IN RETINAL PIGMENT EPITHELIUM

The present invention provides an isolated nucleic acid molecule comprising, or consisting of, the nucleic acid sequence of SEQ ID NO:1 or a nucleic acid sequence of at least 1000 bp having at least 80% identity to said sequence of SEQ ID NO:1, wherein said isolated nucleic acid molecule specifically leads to the expression in cells of the retinal pigment epithelium of a gene when operatively linked to a nucleic acid sequence coding for said gene. 1. An isolated nucleic acid molecule comprising a nucleic acid sequence of at least 1000 base pairs having at least 80% identity to the nucleic acid sequence of SEQ ID NO: 1 , wherein said isolated nucleic acid molecule is effective to promote to the specific expression of a gene in a cell of the retinal pigment epithelium , wherein a nucleic acid sequence coding for said gene is operatively linked to said isolated nucleic acid molecule.2. The isolated nucleic acid molecule of claim 1 , further comprising a minimal promoter-of SEQ ID NO:2.3. (canceled)4. An expression cassette comprising the isolated nucleic acid molecule according to .5. A vector comprising the expression cassette of .6. The vector of claim 5 , wherein said vector is a viral vector.7. (canceled)8. A method of expressing a gene in a cell of the retinal pigment epithelium claim 4 , the method comprising transfecting an isolated cell claim 4 , a cell line claim 4 , or a cell population with the expression cassette according to claim 4 , wherein said transfecting is effective to induce expression of the gene by the isolated cell claim 4 , the cell line claim 4 , or the cell population claim 4 , wherein said cell is a retinal pigment epithelium cell or said cell line or cell population comprises cells of the retinal pigment epithelium.9. An isolated cell comprising the expression cassette of .10. The cell of claim 9 , wherein the expression cassette is stably integrated into the genome of said cell.11. The isolated nucleic acid molecule of claim 1 , wherein ...

Method of Treating or Retarding the Development of Blindness

A method for treating an ocular disorder characterized by the defect or absence of a normal gene in the ocular cells of a human or animal subject involves administering to the subject by subretinal injection an effective amount of a recombinant adeno-associated virus carrying a nucleic acid sequence encoding the normal gene under the control of a promoter sequence which expresses the product of the gene in the ocular cells. The ocular cells are preferably retinal pigment epithelial (RPE) cells, and the gene is preferably an RPE-specific gene, e.g., RPE65. The promoter is one that can express the gene product in the RPE cells. Compositions for subretinal administration are useful in this method. 1. A method for treating an ocular disorder characterized by a defect or absence of a normal gene in ocular cells of a human subject , the method comprising the step ofadministering to said subject by subretinal injection a recombinant adeno-associated virus (rAAV) comprising a nucleic acid sequence encoding a normal gene under the control of a promoter sequence which expresses the product of the gene in ocular cells, wherein expression of the normal gene provides to the cells the product necessary to restore or improve visual function in said subject, wherein said normal gene is the LCA5 or RP12 gene.2. A composition for treatment of an ocular disorder characterized by a defect or absence of a normal gene in ocular cells of a subject , said composition comprising t of a recombinant adeno-associated virus (rAAV) carrying a nucleic acid sequence encoding said normal gene under the control of a promoter sequence which expresses the product of said normal gene in said ocular cells , formulated with a carrier and additional components suitable for subretinal injection ,. wherein said normal gene is the LCA5 or RP12 gene3. The method according to claim 1 , wherein the ocular disorder is Leber congenital amaurosis (LCA) and said normal gene is LCA5.4. The method according to claim ...

REAGENTS AND METHODS FOR MODULATING CONE PHOTORECEPTOR ACTIVITY

The present invention provides reagents and methods for modulating cone photoreceptor activity, and devices for assessment of cone photoreceptor activity.

METHODS OF MAKING PLATELETS COMPRISING MODIFIED RECEPTORS AND USES THEREOF

Disclosed herein are methods of producing platelets comprising a modified receptor, therapeutic agents, peptides, and/or bioactive molecules. The cells produced by the methods disclosed herein can be used to treat, manage, prevent and diagnosis, for example, lysosomal storage diseases, diabetes and cancer. The cells produced by the methods disclosed herein can be engineered to comprise receptors capable of activating platelets to trigger the release of enzymes, biomolecules or therapeutic agents upon binding to specific drugs and/or binding to tissue specific peptides. 1. A nucleic acid construct comprising: a first genetic circuit comprising a tissue-specific promoter operatively linked to a sequence capable of encoding a modified receptor.2. The nucleic acid construct of claim 1 , wherein the tissue-specific promoter is CXCL4 claim 1 , GPIIb claim 1 , or PTPRC.35.-. (canceled)6. The nucleic acid construct of claim 1 , wherein the modified receptor is a modified G-protein coupled receptor (GPCR) or a modified protease-activated receptor (PAR).7. The nucleic acid construct of claim 6 , wherein the modified GPCR is a Gq claim 6 , a Gi claim 6 , a Gs or a G/GGPCR.8. The nucleic acid construct of claim 6 , wherein the modified PAR is PAR1 claim 6 , PAR2 claim 6 , PAR3 or PAR4.9. The nucleic acid construct of claim 1 , further comprising a second genetic circuit claim 1 , wherein the second genetic circuit comprises one or more megakaryocyte differentiation genes.10. The nucleic acid construct of claim 9 , wherein the one or more megakaryocyte differentiation genes are HoxB4 claim 9 , GATA1 claim 9 , c-MYC claim 9 , BMI1 claim 9 , BCL-XL claim 9 , PLK-1 or a combination thereof.1115.-. (canceled)16. The nucleic acid construct of claim 9 , wherein the first genetic circuit or the second genetic circuit further comprises a gene of interest.17. The nucleic acid construct of claim 16 , wherein the gene of interest is a therapeutic agent.18. (canceled)19. (canceled)20. A ...

METHOD AND COMPOSITION TO INCREASE RADIATION-INDUCED TUMOR THERAPEUTIC EFFECTS

Disclosed herein are methods and compositions for treating cancer by increasing radiation-induced damage to cancer without increasing radiation-induced side effects by increasing secretory ASMase levels specifically in tumor endothelium, and inducing apoptosis of tumor endothelial cells by treating the tumor with radiation. ASMase levels are increased in tumor endothelium by administration of a recombinant DNA construct comprising a region coding for a functional ASMase linked to particular transcriptional regulatory sequences that confer tissue-specific expression of ASMase. 120-. (canceled)21. A method for treating cancer in a subject in need thereof comprising(a) administering to the subject an effective amount of an expression vector comprising a recombinant DNA construct comprising a region coding for a functional secretory ASMase linked to at least one transcriptional regulatory sequence that confers tumor endothelium-specific expression of the secretory ASMase, wherein the expression vector is administered intravenously; and(b) exposing the subject to radiation therapy.22. The method of claim 21 , wherein the cancer is a solid tumor.23. The method of claim 21 , wherein the expression vector is a viral expression vector.24. The method of claim 23 , wherein the viral expression vector is replication defective.25. The method of claim 23 , wherein the viral expression vector is an adenovirus vector.26. The method of claim 21 , wherein the at least one transcriptional regulatory sequence drives expression of the secretory ASMase in the angiogenic endothelium of a tumor.27. The method of claim 21 , wherein the at least one transcriptional regulatory sequence is selected from the group consisting of an endothelial-specific promoter and a hypoxia-inducible enhancer.28. The method of claim 27 , wherein the endothelial-specific promoter is pre-proendothelin-1 (PPE-1) promoter or PPE-1 (x3).29. The method of claim 27 , wherein the hypoxia-inducible enhancer is HIF-2α- ...

Constructs containing multiple expression cassettes for cancer therapy

The present invention relates to the field of cancer treatment, particularly to a novel constructs useful for treating tumors expressing H19 and/or IGF-II. More specifically, the invention provides compositions and methods utilizing a nucleic acid construct enabling expression of a cytotoxic gene product directed by more than one tumor specific promoter.

Adeno-associated virus (aav) clades, sequences, vectors containing same, and uses therefor

Sequences of novel adeno-associated virus capsids and vectors and host cells containing these sequences are provided. Also described are methods of using such host cells and vectors in production of rAAV particles. AAV-mediated delivery of therapeutic and immunogenic genes using the vectors of the invention is also provided.

RESCUING VOLTAGE-GATED SODIUM CHANNEL FUNCTION IN INHIBITORY NEURONS

Selectively providing voltage-gated sodium channel function sufficient to rescue impaired Nav1.1 function to inhibitory neurons is described. Provided voltage-gated sodium channel function sufficient to rescue impaired Nav1.1 function in inhibitory neurons can be used to treat disorders such as epilepsy, and more particularly, Dravet Syndrome. 2. The method of claim 1 , wherein the viral vectors comprise SEQ ID NO: 47 and SEQ ID NO: 48; SEQ ID NO: 49 and SEQ ID NO: 50; or SEQ ID NO: 51 and SEQ ID NO: 52.3. The method of claim 1 , wherein the viral vectors comprise SEQ ID NO: 58 claim 1 , SEQ ID NO: 59 claim 1 , SEQ ID NO: 60 claim 1 , and SEQ ID NO: 61.4. An expression construct comprising (i) an enhancer consisting of SEQ ID NO: 3; (ii) a promoter; and (iii) a coding sequence encoding a protein that rescues voltage-gated sodium channel function in a cell or subject in need thereof.5. The expression construct of claim 4 , wherein the coding sequence comprises NavSheP-D60N claim 4 , NavBp claim 4 , NavMs claim 4 , 3×HA-NavSheP-D60N claim 4 , 3×HA-NavBp claim 4 , 3×HA-NavMs claim 4 , His-NavMs claim 4 , human SCN1A claim 4 , mouse Scn1a claim 4 , human SCN1A-3×HA claim 4 , and/or mouse Scn1a-3×HA.6. The expression construct of claim 4 , wherein the promoter comprises minBglobin or minCMV.7. The expression construct of claim 4 , wherein the expression construct is within an adeno-associated viral (AAV) vector.8. The expression construct of claim 4 , wherein the expression construct comprises a coding sequence for a reporter protein.9. The expression construct of claim 8 , wherein the reporter protein comprises a fluorescent reporter protein.10. The expression construct of claim 4 , wherein the expression construct comprises or encodes a skipping element.11. The expression construct of claim 10 , wherein the skipping element comprises a 2A peptide or an internal ribosome entry site (IRES).12. The expression construct of claim 11 , wherein the 2A peptide comprises T2A ...

ADENO-ASSOCIATED VIRUS (AAV) SEROTYPE 8 SEQUENCES, VECTORS CONTAINING SAME, AND USES THEREFOR

Sequences of a serotype 8 adeno-associated virus and vectors and host cells containing these sequences are provided. Also described are methods of using such host cells and vectors in production of rAAV particles. 1. A recombinant nucleic acid molecule:(a) encoding an AAV8 vp1 capsid protein having a sequence comprising amino acids 1 to 738 of SEQ ID NO: 2; or(b) comprising nucleotides 2121 to 4334 of SEQ ID NO: 1, or a nucleotide sequence at least 99% identical to nucleotides 2121 to 4334 of SEQ ID NO: 1,wherein the recombinant nucleic acid molecule does not contain an AAV inverted terminal repeat.2. The recombinant nucleic acid molecule according to claim 1 , which further comprises a functional rep gene.3. The recombinant nucleic acid molecule according to claim 1 , wherein said recombinant nucleic acid molecule comprises a sequence selected from:vp1, nt 2121 to 4334;vp2, nt 2532 to 4334; orvp3, nt 2730 to 4334 of SEQ ID NO: 1.4. The recombinant nucleic acid molecule according to claim 1 , wherein said recombinant nucleic acid molecule is a plasmid.5. A host cell transfected with the recombinant nucleic acid molecule according to .6. The host cell according to claim 5 , which further comprises a functional rep gene claim 5 , a minigene comprising AAV inverted terminal repeats (ITRs) and a transgene claim 5 , and sufficient helper functions to permit packaging of the minigene into the AAV capsid.7. The host cell according to claim 6 , which further comprises a nucleic acid sequence '(b) comprising nucleotides 2532 to 4334 of SEQ ID NO: 1, or a nucleotide sequence at least 99% identical to nucleotides 2532 to 4334 of SEQ ID NO: 1.', '(a) encoding an AAV8 vp2 capsid protein having a sequence comprising amino acids 138 to 738 of SEQ ID NO: 2; or'}8. The host cell according to claim 6 , which further comprises a nucleic acid sequence(a) encoding an AAV8 vp3 capsid protein having a sequence comprising amino acids 204 to 738 of SEQ ID NO: 2; or(b) comprising nucleotides ...

ADENO-ASSOCIATED VIRUS VECTORS FOR TREATMENT OF GLYCOGEN STORAGE DISEASE

The present disclosure describes improved adeno-associated virus (AAV) vectors for gene therapy applications in the treatment of glycogen storage disease, particularly glycogen storage disease type Ia (GSD-Ia). Described are recombinant nucleic acid molecules, vectors and recombinant AAV that include a G6PC promoter/enhancer, a synthetic intron, a G6PC coding sequence (such as a wild-type or codon-optimized G6PC coding sequence), and stuffer nucleic acid sequence situated between the G6PC promoter/enhancer and the intron, as well as between the intron and the G6PC coding sequence. The recombinant AAVs disclosed herein exhibit highly efficient liver transduction and are capable of correcting metabolic abnormalities in an animal model of GSD-Ia. 1. A recombinant nucleic acid molecule comprising a G6PC promoter/enhancer , a synthetic intron , and a G6PC coding region , wherein the nucleotide sequence of the recombinant nucleic acid molecule is at least 90% identical to nucleotides 182-4441 of SEQ ID NO: 1 , and wherein the recombinant nucleic acid comprises is at least one nucleotide substitution in the G6PC coding region that results in a coding change at residue 3 , 54 , 139 , 196 , 199 , 242 , 247 , 292 , 298 , 301 , 318 , 324 , 332 , 347 , 349 , 350 or 353 of the human G6PC protein of SEQ ID NO: 4.2. The recombinant nucleic acid molecule of claim 1 , wherein the nucleotide substitution in the G6PC coding region results in a coding change at residue 298 of the human G6PC protein of SEQ ID NO: 4.3. The recombinant nucleic acid molecule of claim 1 , further comprising 5′ and 3′ inverted terminal repeat (ITR) sequences claim 1 , wherein the recombinant nucleic acid molecule comprises a nucleotide sequence at least 90% identical to nucleotides 17-4819 of SEQ ID NO: 1.4. The recombinant nucleic acid molecule of claim 1 , comprising a nucleotide sequence at least 90% identical to SEQ ID NO: 1.5. A vector comprising the recombinant nucleic acid molecule of .6. The vector ...

A MODIFIED MGLUR6 PROMOTER AND METHODS OF USE

The invention provides nucleic acids and nucleic acid expression vectors containing optimized mGluR6 promoters for expression of transgenes in the retina. The compositions and methods of the invention are useful for expression of gene products to preserve, improve, or restore phototransduction or vision. 1. An isolated nucleic acid molecule comprisinga. an mGluR6 enhancer or a variant thereof;b. an mGluR6 promoter or a variant thereof; andc. optionally, an intron 4 of the mGluR6 gene or a variant thereof; andd. optionally, an intron 3 of the mGluR6 gene or a variant thereof.2. The isolated nucleic acid molecule of claim 1 , whereina. said mGluR6 enhancer variant is at least 70% identical to SEQ ID NO: 1 or SEQ ID NO: 2;b. said mGluR6 promoter variant is at least 70% identical to SEQ ID NO: 3, SEQ ID NO: 4, SEQ ID NO: 5, or SEQ ID NO: 6;c. said intron 4 of the mGluR6 gene variant is at least 70% identical to SEQ ID NO: 7 or SEQ ID NO: 8; ord. said intron 3 of the mGluR6 gene variant is at least 70% identical to SEQ ID NO: 9 or SEQ ID NO: 10.3. The isolated nucleic acid molecule of claim 1 , wherein said mGluR6 enhancer comprises the nucleic acid sequence of SEQ ID NO: 1 or SEQ ID NO: 2 claim 1 , wherein said mGluR6 promoter comprises the nucleic acid sequence of SEQ ID NO: 3 claim 1 , SEQ ID NO: 4 claim 1 , SEQ ID NO: 5 claim 1 , or SEQ ID NO: 6 claim 1 , wherein said intron 4 of the mGluR6 gene comprises the nucleic acid sequence of SEQ ID NO: 7 or SEQ ID NO: 8 claim 1 , and wherein said intron 3 of the mGluR6 gene comprises the nucleic acid sequence of SEQ ID NO: 9 or SEQ ID NO: 10.4. The isolated nucleic acid molecule of any one of the preceding claims claim 1 , wherein the intron 4 of the mGluR6 gene is upstream of the intron 3 of the mGluR6 gene claim 1 , wherein the intron 3 of the mGluR6 gene is upstream of the mGluR6 enhancer claim 1 , and wherein the mGluR6 enhancer is upstream of the mGluR6 promoter.5. A nucleic acid expression vector comprising a promoter ...

ADENO-ASSOCIATED VIRUS (AAV) CLADES, SEQUENCES, VECTORS CONTAINING SAME, AND USES THEREFOR

Sequences of novel adeno-associated virus capsids and vectors and host cells containing these sequences are provided. Also described are methods of using such host cells and vectors in production of rAAV particles. AAV-mediated delivery of therapeutic and immunogenic genes using the vectors of the invention is also provided. 1. A recombinant nucleic acid molecule:(a) encoding an AAVhu37 vp1 capsid protein having a sequence comprising amino acids 1 to 738 of SEQ ID NO: 88; or(b) comprising nucleotides 1 to 2214 of SEQ ID NO: 10, or a nucleotide sequence at least 99% identical to nucleotides 1 to 2214 of SEQ ID NO: 10,wherein the recombinant nucleic acid molecule does not contain an AAV inverted terminal repeat.2. The recombinant nucleic acid molecule according to claim 1 , which further comprises a functional rep gene.3. The recombinant nucleic acid molecule according to claim 1 , wherein said recombinant nucleic acid molecule comprises a sequence selected from:vp1 , nt 1 to 2214;vp2, nt 412 to 2214; orvp3, nt 610 to 2214 of SEQ ID NO: 10.4. The recombinant nucleic acid molecule according to claim 1 , wherein said recombinant nucleic acid molecule is a plasmid.5. A host cell transfected with the recombinant nucleic acid molecule according to .6. The host cell according to claim 5 , which further comprises a functional rep gene claim 5 , a minigene comprising AAV inverted terminal repeats (ITRs) and a transgene claim 5 , and sufficient helper functions to permit packaging of the minigene into the AAV capsid.7. The host cell according to claim 6 , which further comprises a nucleic acid sequence(a) encoding an AAVhu37 vp2 capsid protein having a sequence comprising amino acids 138 to 738 of SEQ ID NO: 88; or(b) comprising nucleotides 412 to 2214 of SEQ ID NO: 10, or a nucleotide sequence at least 99% identical to nucleotides 412 to 2214 of SEQ ID NO: 10.8. The host cell according to claim 6 , which further comprises a nucleic acid sequence(a) encoding an AAVhu37 vp3 ...

RECOMBINANT AAV VARIANTS AND USES THEREOF

The disclosure in some aspects relates to recombinant adeno-associated viruses having distinct tissue targeting capabilities. In some aspects, the disclosure relates to gene transfer methods using the recombinant adeno-associated viruses. In some aspects, the disclosure relates to isolated AAV capsid proteins and isolated nucleic acids encoding the same. 1. A recombinant expression vector comprising a sequence selected from the group consisting of: SEQ ID NO: 1 to 47 , or a fragment thereof that does not encode a peptide that is identical to a sequence of any one of SEQ ID NOs: 98 to 100.2. An isolated AAV capsid protein comprising an amino acid sequence selected from the group consisting of: SEQ ID NOs: 51 to 97.3. An isolated AAV capsid protein comprising a sequence selected from the group consisting of: SEQ ID NOs: 51 to 61 , wherein an amino acid of the sequence that is not identical to a corresponding amino acid of the sequence set forth as SEQ ID NO: 98 is replaced with a conservative substitution.4. An isolated AAV capsid protein comprising a sequence selected from the group consisting of: SEQ ID NOs: 62 to 67 , wherein an amino acid of the sequence that is not identical to a corresponding amino acid of the sequence set forth as SEQ ID NO: 99 is replaced with a conservative substitution.5. An isolated AAV capsid protein comprising a sequence selected from the group consisting of: SEQ ID NOs: 68 to 97 , wherein an amino acid of the sequence that is not identical to a corresponding amino acid of the sequence set forth as SEQ ID NO: 100 is replaced with a conservative substitution.6. A peptide fragment of the isolated AAV capsid protein of any one of to that is not identical to a sequence of any one of SEQ ID NOs: 98 to 100.7. An isolated AAV capsid protein comprising the peptide fragment of .8. An recombinant expression vector comprising a nucleic acid sequence encoding the isolated AAV capsid protein of any one of to .9. A composition comprising the isolated ...

Method of detecting and/or identifying adeno-associated virus (aav) sequences and isolating novel sequences identified thereby

Adeno-associated virus rh.20 sequences, vectors containing same, and methods of use are provided.

Mito-Ob: A Transgenic Mouse Model for Obesity

An obese mouse model was developed by overexpressing the mitochondrial protein prohibitin (PHB) in white adipose tissue (WAT) specific manner driven by adipocyte protein 2 (aP2) promoter. These mice begin to develop obesity as a result of mitochondrial remodeling (upregulation of mitochondrial biogenesis and function) in WAT.

RECOMBINANT AAV VARIANTS AND USES THEREOF

The disclosure in some aspects relates to recombinant adeno-associated viruses having distinct tissue targeting capabilities. In some aspects, the disclosure relates to gene transfer methods using the recombinant adeno-associated viruses. In some aspects, the disclosure relates to isolated AAV capsid proteins and isolated nucleic acids encoding the same. 142.-. (canceled)43. A recombinant adeno-associated viral (rAAV) particle comprising an AAV capsid and at least one transgene , wherein the AAV capsid comprises a protein comprising an amino acid sequence selected from any one of SEQ ID NOs: 79 to 87.44. A composition comprising the rAAV particle of claim 43 , and a pharmaceutically acceptable carrier.45. A method for delivering a transgene to a subject comprising administering the rAAV particle of to the subject claim 43 , wherein the rAAV particle infects cells of a target tissue of the subject.46. The method of claim 45 , wherein the at least one transgene is a protein coding gene.47. The method of claim 45 , wherein the at least one transgene encodes a small interfering nucleic acid selected from a miRNA or an shRNA.48. The method of claim 45 , wherein the target tissue is skeletal muscle claim 45 , heart claim 45 , liver claim 45 , pancreas claim 45 , spleen claim 45 , brain claim 45 , or lung.49. The method of claim 45 , wherein the target tissue is heart tissue.50. The method of claim 45 , wherein the rAAV particle is administered intravenously claim 45 , transdermally claim 45 , intraocularly claim 45 , intrathecally claim 45 , orally claim 45 , intramuscularly claim 45 , subcutaneously claim 45 , intranasally claim 45 , or by inhalation.51. A method for generating a somatic transgenic non-human animal model comprising administering the rAAV particle of to a non-human animal claim 43 , wherein the rAAV particle infects cells of a target tissue of the non-human animal.52. A somatic transgenic non-human animal model produced by the method of .53. A recombinant ...

ANIMAL MODELS OF AGE-RELATED DISORDERS AND AGE-SENSITIVE TRAITS ASSOCIATED WITH SENESCENCE-INDUCING STIMULI AND USES THEREOF

This disclosure provides non-human animal models for age-related disorders and age-sensitive traits, particularly those caused by senescence-inducing stimuli, wherein the models comprise transgenes selectively expressed by senescent cells. The disclosure further provides methods for identifying therapeutic agents effective for treating or preventing age-related disorders and age-sensitive traits using the animal models, therapeutic agents identified using such methods, pharmaceutical compositions comprising the identified therapeutic agents, and methods of treating or preventing age-related disorders and age-sensitive traits. 1. A non-human animal model for aging comprising a non-human animal that (a) exhibits an age-related disorder or age-sensitive trait , and (b) comprises a transgene selectively expressed by senescent cells , wherein the animal is exposed to or treated with a senescence-inducing stimulus.2. The animal model of claim 1 , wherein the transgene comprises a senescent cell-specific promoter.3. The animal model of claim 2 , wherein the senescent cell-specific promoter is derived from p16.4. The animal model of claim 1 , wherein the transgene expresses at least one detectable label claim 1 , a cytotoxic agent claim 1 , a cytotoxicity-activating molecule claim 1 , an RNA claim 1 , or a combination thereof.5. The animal model of claim 4 , wherein the detectable label is selected from the group consisting of (a) luciferase; (b) a red fluorescent protein; (c) a green fluorescent protein; and (d) a luciferase and a red fluorescent protein.6. The animal model of claim 4 , wherein the cytotoxicity-activating molecule is a truncated herpes simplex virus thymidine kinase or a FK506-binding protein (FKBP)-caspase fusion polypeptide.7. (canceled)8. The animal model of claim 1 , wherein the transgene comprises (a) a p16promoter operatively linked to a polynucleotide sequence encoding a FKBP-caspase fusion polypeptide (p16-FKBP-caspase transgene) claim 1 , and to a ...

Adeno-associated virus (aav) clades, sequences, vectors containing same, and uses therefor

Sequences of novel adeno-associated virus capsids and vectors and host cells containing these sequences are provided. Also described are methods of using such host cells and vectors in production of rAAV particles. AAV-mediated delivery of therapeutic and immunogenic genes using the vectors of the invention is also provided.

S100b mini-promoters

Isolated polynucleotides comprising a S100B mini-promoters are provided. The mini-promoter may be operably linked to an expressible sequence, e.g. reporter genes, genes encoding a polypeptide of interest, regulatory RNA sequences such as miRNA, siRNA, anti-sense RNA, etc., and the like. In some embodiments a cell comprising a stable integrant of an expression vector is provided, which may be integrated in the genome of the cell. The promoter may also be provided in a vector, for example in combination with an expressible sequence. The polynucleotides find use in a method of expressing a sequence of interest, e.g. for identifying or labeling cells, monitoring or tracking the expression of cells, gene therapy, etc.

GENE THERAPY TO IMPROVE VISION

The invention relates to the use of gene therapy vectors to improve vision by introducing into healthy rod photoreceptor cells of a patient suffering from cone photoreceptor dysfunction and/or degeneration a nucleic acid encoding a gene product that is light-sensitive and/or that modulates endogenous light-sensitive signaling in a photoreceptor cell, such that the range of light intensities to which the rod photoreceptor responds is extended and/or the speed at which the rod photoreceptor responds to light is increased. 1. A vector comprising a nucleic acid encoding a gene product that is light-sensitive and/or that modulates endogenous light-sensitive signaling in a photoreceptor cell , for use in a method of improving vision in a patient with cone photoreceptor dysfunction and/or degeneration by introduction of said nucleic acid into healthy rod photoreceptors in the retina of the patient and expression of said gene product therein , such that the range of light intensities to which the rod photoreceptor responds is extended and/or the speed at which the rod photoreceptor responds to light is increased.2. A vector for use according to wherein the nucleic acid encodes a protein that changes membrane conductance in a way that results in rod hyperpolarisation (outward current flow) upon light stimulation.3. A vector for use according to wherein the nucleic acid encodes (a) a light-sensitive or light-gated G-coupled membrane protein claim 2 , ion channel claim 2 , ion pump or ion transporter (b) a member of the RGS9 complex claim 2 , or (c) another protein that increases the speed of the endogenous rod signaling mechanism.4. A vector for use according to wherein the light-gated molecule is ArchT claim 3 , Jaws (cruxhalorhodopsin) claim 3 , iC1C2 claim 3 , or the member of the RGS9 complex is R9AP.5. A vector for use according to any one of the preceding claims which is a viral vector.6. A vector for use according to which is an adeno associated virus (AAV) vector.7. ...

METHOD OF DETECTING AND/OR IDENTIFYING ADENO-ASSOCIATED VIRUS (AAV) SEQUENCES AND ISOLATING NOVEL SEQUENCES IDENTIFIED THEREBY

Adeno-associated virus rh.10 sequences, vectors containing same, and methods of use are provided. 1. A method for delivering a transgene product to a subject , said method comprising administering an adeno-associated virus (AAV) comprising an AAV capsid comprising vp1 , vp2 and vp3 proteins , said vp3 having the amino acid sequence of 204 to 738 of SEQ ID NO: 81 or an AAV vp3 protein having a sequence at least 95% identical to the full length amino acid sequence of 204 to 738 of SEQ ID NO: 81 , said AAV having packaged in the capsid a nucleic acid molecule comprising at least one AAV inverted terminal repeat (ITR) and a non-AAV nucleic acid sequence which encodes a gene product operably linked to sequences which direct expression thereof in a host cell.2. The method according to claim 1 , wherein the AAVrh10 capsid comprises vp1 proteins having an amino acid sequence of about amino acids 1 to 738 of SEQ ID NO:81.3. The method according to claim 1 , wherein the sequence is at least 97% identical to the vp1 and/or vp3 of SEQ ID NO: 814. The method according to claim 1 , wherein the sequence is at least 99% identical to the vp1 and/or vp3 of SEQ ID NO: 81.5. The method according to claim 1 , wherein the vp2 protein has an amino acid sequence of about amino acids 138 to 738 of SEQ ID NO:81.6. The method according to claim 1 , wherein the gene product is a vascular endothelial growth factor (VEGF).7. The method according to claim 1 , wherein the gene product is selected from β-glucuronidase (GUSB) and alpha-1 antitrypsin (A1AT).8. The method according to claim 1 , wherein the gene product is a factor IX protein.9. The method according to claim 1 , wherein the gene product is a factor VIII protein.10. The method according to claim 1 , wherein the gene product is erythropoietin.11. An isolated capsid protein comprising an AAVrh10 protein selected from the group consisting of:vp1 capsid protein, amino acids (aa) 1 to 738 of SEQ ID NO: 81;vp2 capsid protein, aa 138 to 738 of ...

DELIVERY OF POLYNUCLEOTIDES USING RECOMBINANT AAV9

The present invention relates to Adeno-associated virus 9 methods and materials useful for systemically delivering polynucleotides across the blood brain barrier. Accordingly, the present invention also relates to methods and materials useful for systemically delivering polynucleotides to the central and peripheral nervous systems. The present invention also relates to Adeno-associated virus type 9 methods and materials useful for intrathecal delivery of polynucleotides. Use of the methods and materials is indicated, for example, for treatment of lower motor neuron diseases such as spinal muscle atrophy and amyotrophic lateral sclerosis as well as Pompe disease and lysosomal storage disorders. Use of the methods and materials is also indicated, for example, for treatment of Rett syndrome. 1. A method of delivering a polynucleotide encoding a therapeutic peptide for the treatment of mucopolysaccharidosis IIIA (MPS IIIA) to the central nervous system of a patient suffering from MPS IIIA , the method comprising administering a recombinant adeno-associated virus 9 (rAAV9) to the patient by direct intravenous injection , wherein the rAAV9 comprises the polynucleotide in a self-complementary genome.2. A method of treating mucopolysaccharidosis IIIA (MPS IIIA) in a patient suffering from MPS IIIA , the method comprising administering a rAAV9 to the patient by direct intravenous injection , wherein the rAAV9 comprises the polynucleotide in a self-complementary genome.3. The method of or wherein the rAAV9 is delivered across the blood brain barrier (BBB) to the patient's central nervous system.4. The method of wherein the rAAV9 is delivered to the patient's brain.5. The method of wherein the rAAV9 is delivered to the patient's spinal cord.6. A method of delivering a polynucleotide encoding a therapeutic peptide for the treatment of mucopolysaccharidosis IIIB (MPS IIIB) to the central nervous system of a patient suffering from MPS IIIB claim 3 , the method comprising ...

Gene Therapy for the Treatment of CNGB1-linked Retinitis Pigmentosa

The present invention relates to a polynucleotide comprising a promoter comprising a human photoreceptor-specific promoter element, a core promoter and at least one transgene. Further, the invention provides a plasmid comprising the polynucleotide, a viral vector comprising the polynucleotide and a pharmaceutical composition comprising the polynucleotide. The invention also relates to the plasmid, the viral vector or the pharmaceutical composition for use as a medicament, in particular for use in the therapy of diseases of the retina.

TARGETED DISRUPTION OF A CSF1-DAP12 PATHWAY MEMBER GENE FOR THE TREATMENT OF NEUROPATHIC PAIN

The invention provides compositions and methods for treating neuropathic pain. Specifically, the disclosure provides a polynucleotide comprising a trigeminal ganglion (TGG) or dorsal root ganglion (DRG) promoter operably linked to a recombinant nucleic acid encoding an endonuclease that binds to a nucleotide sequence in the human colony stimulating factor 1 (hCSF1) gene and a method of using the polynucleotide or a vector comprising the polynucleotide for treatment of neuropathic pain. 1. A polynucleotide comprising a trigeminal ganglion (TGG) or dorsal root ganglion (DRG) promoter operably linked to a recombinant nucleic acid encoding an endonuclease that binds to a nucleotide sequence in the human colony stimulating factor 1 (hCSF1) gene.2. The polynucleotide of claim 1 , wherein binding of the endonuclease to the nucleotide sequence decreases claim 1 , reduces claim 1 , or eliminates hCSF gene expression in a dorsal root ganglion cell.3. The polynucleotide of claim 1 , wherein the TGG or DRG promoter is selected from the group consisting of: an hSYN1 promoter claim 1 , a TRPV1 promoter claim 1 , a Nav1.7 promoter claim 1 , a Nav1.8 promoter claim 1 , a Nav1.9 promoter claim 1 , a CAG promoter claim 1 , and an Advillin promoter.4. The polynucleotide of any of - claim 1 , wherein the nucleotide sequence in the hCSF1 gene is selected from the group consisting of: an hCSF1 gene regulatory region claim 1 , an hCSF1 promoter claim 1 , an hCSF1 transcription start site claim 1 , an hCSF1 exon sequence claim 1 , an hCSF1 intronic sequence claim 1 , and an hCSF1 5′ or 3′ untranslated region.5. The polynucleotide of claim 1 , wherein the endonuclease is an endonuclease that is engineered to bind the nucleotide sequence of the hCSF1 gene.6. The polynucleotide of claim 5 , wherein the engineered endonuclease is a homing endonuclease claim 5 , a transcription activator-like effector nucleases (TALENs) claim 5 , a zinc finger nuclease (ZFN) claim 5 , a Type II clustered ...

GENE THERAPY FOR JUVENILE BATTEN DISEASE

Compositions and methods for the treatment of Juvenile Neuronal Ceroid Lipofuscinosis (JNCL), also known as Juvenile Batten Disease, are provided herein. In certain embodiments the compositions include but are not limited to adeno-associated viral (AAV) constructs, including self-complementary adeno-associated viral (sc-AAV) constructs, that express the human gene CLN3 (or a CLN3 cDNA). 1. A method for treating or preventing Juvenile neuronal ceroid lipofuscinosis (JNCL) in a mammal , the method comprising administering to the mammal a composition comprising a cell or a population of cells transformed with a recombinant vector comprising:(i) recombinant adeno-associated virus (AAV) genome or a derivative thereof, wherein the AAV is selected from AAV1, AAV2, AAV4, AAV5, AAV6, AAV8, or AAV9; and(ii) a promoter that drives low expression of CLN3 operably linked to a polynucleotide sequence encoding CLN3 or an equivalent thereof, wherein the CLN3 is expressed at an effective amount.2. The method of claim 1 , wherein the transformed cell or population of cells comprises a cell or tissue of the central nervous system (CNS) of the mammal.3. (canceled)4. The method of claim 1 , wherein the mammal is selected from the group of a human; a human neonate claim 1 , a human infant claim 1 , or a human adolescent; a human adult; asymptomatic for JNCL; or symptomatic for JNCL.5. The method of claim 4 , wherein the human is homozygous for a CLN3 mutation.69.-. (canceled)10. The method of claim 4 , wherein the symptomatic mammal for JNCL presents with: one or more of blindness; seizures; motor loss; or cognitive decline.1113.-. (canceled)14. The method of claim 1 , wherein the CLN3 is a human CLN3.15. (canceled)16. The method of claim 1 , wherein the AAV genome is from a naturally derived serotype claim 1 , an isolate claim 1 , or a clade of AAV.17. (canceled)18. The method of claim 1 , wherein the AAV serotype is AAV9 or AAV2.19. (canceled)20. The method of claim 1 , wherein the ...

NOVEL MICRO-DYSTROPHINS AND RELATED METHODS OF USE

Nucleotide sequences including a micro-dystrophin gene are provided. The micro-dystrophin genes may be operatively linked to a regulatory cassette. Methods of treating a subject having, or at risk of developing, muscular dystrophy, sarcopenia, heart disease, or cachexia are also provided. The methods may include administering a pharmaceutical composition including the micro-dystrophin gene and a delivery vehicle to a subject. Further, the methods may include administering the pharmaceutical composition a subject having Duchenne muscular dystrophy or Becker muscular dystrophy. 1. A method of treating a subject having a muscular dystrophy , the method comprising:administering to the subject a therapeutically effective amount of a pharmaceutical composition comprising:a micro-dystrophin gene encoding a protein comprising:an amino-terminal actin-binding domain;a β-dystroglycan binding domain; anda spectrin-like repeat domain consisting of five spectrin-like repeats, including spectrin-like repeat 1 (SR1), spectrin-like repeat 16 (SR16), spectrin-like repeat 17 (SR17), spectrin-like repeat 23 (SR23), and spectrin-like repeat 24 (SR24);wherein the micro-dystrophin gene is operatively linked to a regulatory cassette; and a delivery vehicle.2. The method of claim 1 , wherein the protein encoded by the micro-dystrophin gene further comprises at least a portion of a hinge domain.3. The method of claim 2 , wherein the hinge domain is selected from at least one of a Hinge 1 domain claim 2 , a Hinge 2 domain claim 2 , a Hinge 3 domain claim 2 , and a Hinge 4 domain.4. The method of claim 1 , wherein the regulatory cassette is selected from the group consisting of a CK8 promoter and a cardiac troponin T (cTnT) promoter.5. The method of claim 4 , wherein the regulatory cassette is configured to express the micro-dystrophin gene such that a level of expression of the micro-dystrophin gene is at least about 100-fold higher in striated muscle cells than the level of expression of the ...

DOPAMINE RECEPTOR TYPE 2 SPECIFIC PROMOTER AND METHODS OF USE THEREOF

A nucleic acid containing a dopamine receptor type 2-specific promoter (D2SP) is provided. In certain embodiments, the nucleic acid includes a dopamine receptor type 2-specific promoter (D2SP), wherein the D2SP does not include exon 1 of a D2 receptor gene, wherein the D2SP comprises a Kozak sequence, and wherein the D2SP includes a nucleotide sequence having at least 95% sequence identity to the nucleotide sequence set forth in SEQ ID NO: 1. Also provided are expression vectors, genetically modified host cells and kits that include the subject nucleic acid. 1. A nucleic acid comprising a dopamine receptor type 2-specific promoter (D2SP) , wherein the D2SP does not include exon 1 of a D2 receptor gene , wherein the D2SP contains a Kozak sequence , and wherein the D2SP contains a nucleotide sequence having at least 95% sequence identity to the nucleotide sequence set forth in SEQ ID NO: 1.2. The nucleic acid of claim 1 , wherein the Kozak sequence is at the 3′ terminus of the D2SP.3. The nucleic acid of any of and claim 1 , wherein the D2SP contains a BamHI restriction site.4. The nucleic acid of claim 3 , wherein the BamHI restriction site is located 5′ of the Kozak sequence.5. The nucleic acid of any of to claim 3 , wherein the D2SP contains a nucleotide sequence having at least 98% sequence identity to the nucleotide sequence set forth in SEQ ID NO: 1.6. The nucleic acid of any of to claim 3 , wherein the D2SP is operably linked to a nucleotide sequence encoding a gene product that provides a detectable signal.7. The nucleic acid of claim 6 , wherein the gene product that provides a detectable signal is a fluorescent protein.8. The nucleic acid of claim 7 , wherein the fluorescent protein is selected from the group consisting of a green fluorescent protein claim 7 , a yellow fluorescent protein claim 7 , a cyan fluorescent protein claim 7 , a calcium indicator and a voltage indicator.9. The nucleic acid of any of to claim 7 , wherein the D2SP is operably linked to ...

AAV-MEDIATED GENE THERAPY FOR RPGR X-LINKED RETINAL DEGENERATION

Described herein are methods of preventing, arresting progression of or ameliorating vision loss and other conditions associated with retinitis pigmentosa and x-linked retinitis pigmentosa in a subject. The methods include administering to said subject an effective concentration of a composition comprising a recombinant adeno-associated virus (AAV) carrying a nucleic acid sequence encoding a normal retinitis pigmentosa GTPase regulator (RPGR gene), or fragment thereof, under the control of regulatory sequences which express the product of the gene in the photoreceptor cells of the subject, and a pharmaceutically acceptable carrier. 1. A recombinant adeno-associated virus (AAV) carrying a nucleic acid sequence encoding a normal RPGR gene , or fragment thereof , under the control of regulatory sequences which express the product of said gene in the photoreceptor cells of said subject.2. A method of preventing , arresting progression of or ameliorating vision loss associated with retinitis pigmentosa in a subject , said method comprising administering to said subject an effective concentration of a composition comprising a recombinant adeno-associated virus (AAV) carrying a nucleic acid sequence encoding a normal RPGR gene , or fragment thereof , under the control of regulatory sequences which express the product of said gene in the photoreceptor cells of said subject , and a pharmaceutically acceptable carrier. This application is a continuation of U.S. patent application Ser. No. 14/413,884, filed Jan. 9, 2015, which is a US national phase of International Patent Application No. PCT/US2013/022628, filed Jan. 23, 2013, which claims the benefit of the priority of U.S. Provisional Patent Application No. 61/670,355, filed Jul. 11, 2012, now expired. These priority applications are incorporated by reference herein.This invention was made with government support under grant/contract numbers 5R01EY017549, 5R01EY006855, 1P30EY021721, 5R01EY007961, P30EY001583 and ...

DELIVERY OF POLYNUCLEOTIDES USING RECOMBINANT AAV9

The present invention relates to Adeno-associated virus 9 methods and materials useful for systemically delivering polynucleotides across the blood brain barrier. Accordingly, the present invention also relates to methods and materials useful for systemically delivering polynucleotides to the central and peripheral nervous systems. The present invention also relates to Adeno-associated virus type 9 methods and materials useful for intrathecal delivery of polynucleotides. Use of the methods and materials is indicated, for example, for treatment of lower motor neuron diseases such as spinal muscle atrophy and amyotrophic lateral sclerosis as well as Pompe disease and lysosomal storage disorders. Use of the methods and materials is also indicated, for example, for treatment of Rett syndrome. 120-. (canceled)21. A method of treating Rett syndrome in a patient comprising the step of intrathecal administration of an effective dose of a recombinant AAV9 (rAAV9) to the patient , wherein the rAAV9 comprises a methyl-CpG-binding protein 2 (MECP2) polynucleotide in a self-complementary genome.22. The method of wherein the patient is a female patient.23. The method of wherein the dose of rAAV9 administered is 1×10vg/kg or more.24. The method of wherein the MECP2 polynucleotide is expressed in neuronal and glial cells in the central nervous system of the patent.25. The method of wherein the treatment reduces patient seizures.26. The method of wherein the treatment improves patient mobility. The present application is a continuation-in-part of U.S. patent application Ser. No. 13/270,840 filed Oct. 11, 2011. U.S. patent application Ser. No. 13/270,840 is a continuation of U.S. patent application Ser. No. 13/035,777 filed Feb. 25, 2011. U.S. patent application Ser. No. 13/035,777 claims the benefit of priority of U.S. Provisional Application No. 61/308,884, filed Feb. 26, 2010, and is also a continuation-in-part of International Patent Application No. PCT/US09/68818, filed Dec. 18, ...

PHARMACEUTICAL COMPOSITION COMPRISING NANOG SHRNA, AND METHOD OF USING NANOG SHRNA TO TREAT CANCER

The present description relates to an inhibitory RNA molecule, comprising an oligonucleotide that selectively knocks down expression a Nanog pseudogene expressed in many human cancers, a replicating viral vector capable of encoding such inhibitory RNA molecule, pharmaceutical compositions comprising said vector, and methods of treating cancer by administration of said pharmaceutical composition. 1. An inhibitory RNA molecule , comprising an oligonucleotide that knocks down expression of NANOGP8 wherein the oligonucleotide is 15 to 29 nucleotides in length and comprises a sequence selected from the group consisting ofSEQ ID NO:3 (5′-GAGGCAGCAGAGACCGCTGCATGCACTTCCAGCCA-3′); andSEQ ID NO:8 (5′-TCTGACAGGAAGTGGCTGGAAGTGCATGCAG-3′).2. The inhibitory RNA molecule of claim 1 , wherein the inhibitory RNA molecule is selected from the group consisting of SEQ ID NO:54 (CTGCATGCACTTCCAGCCA) claim 1 , SEQ ID NO:55 (TGGCTGGAAGTGCATGCAG) claim 1 , SEQ ID NO:56 (CTGCATGCACTTCCAGCCG) and SEQ ID NO: 57 (TGGCTGGAAGTGCATGCAG).3. The inhibitory RNA molecule of claim 1 , wherein the inhibitory RNA molecule is a double stranded siRNA comprising SEQ ID NO:54 (CTGCATGCACTTCCAGCCA) and SEQ ID NO:55 (TGGCTGGAAGTGCATGCAG) or SEQ ID NO:56 (CTGCATGCACTTCCAGCCG) and SEQ ID NO: 57 (TGGCTGGAAGTGCATGCAG).4. The inhibitory RNA molecule of claim 1 , wherein the inhibitory RNA molecule comprises one or more nucleotide analogs or modifications.5. The inhibitory RNA molecule of claim 1 , wherein the inhibitory RNA molecule is encapsulated in a lipid particle.6. A conditionally replicating viral vector claim 1 , comprising an inhibitory RNA molecule comprising an oligonucleotide that knocks down expression of NANOGP8 wherein the oligonucleotide is 15 to 29 nucleotides in length and comprises a sequence selected from the group consisting ofSEQ ID NO:3 (5′-GAGGCAGCAGAGACCGCTGCATGCACTTCCAGCCA-3′); andSEQ ID NO:8 (5′-TCTGACAGGAAGTGGCTGGAAGTGCATGCAG-3′).7. The conditionally replicating viral vector of claim 6 ...

THERAPEUTIC ADENO-ASSOCIATED VIRUS FOR TREATING POMPE DISEASE

Recombinant AAV (rAAV) vectors comprising a rAVV genome comprising a heterologous nucleic acid encoding a signal peptide and optionally a IGF-2 sequence, fused to an acid alpha-glucosidase (GAA) polypeptide, enabling the GAA polypeptide to be secreted from the liver and targeted to the lysosomes. Particular embodiments relate to a recombinant AAV (rAAV) vector encoding an alpha-glucosidase (GAA) polypeptide, having a liver secretory signal peptide and a targeting IGF2 sequence that binds human cation-independent mannose-6-phosphate receptor (CI-MPR) or to the IGF2 receptor, permitting proper subcellular localization of the GAA polypeptide to lysosomes. Also encompassed are cells, and methods to treat a glycogen storage disease type II (GSD II) disease and/or Pompe Disease with the rAAV vector. 17.-. (canceled)8. The recombinant AAV vector of claim 34 , wherein the promoter is constitutive claim 34 , cell specific or inducible.931.-. (canceled)32. The recombinant AAV vector of claim 34 , wherein the AAV3b serotype comprises one or mutations in a capsid protein selected from any of: 265D claim 34 , 549A claim 34 , Q263Y33. The recombinant AAV vector of claim 34 , wherein the AAV3b serotype is selected from any of: AAV3b265D claim 34 , AAV3b265D549A claim 34 , AAV3b549A or AAV3bQ263Y claim 34 , or AAV3bSASTG.34. A recombinant adenovirus associated (AAV) vector comprising in its genome:a. 5′ and 3′ AAV inverted terminal repeats (ITR) sequences, andb. located between the 5′ and 3′ ITRs, a heterologous nucleic acid sequence encoding a polypeptide comprising an alpha-glucosidase (GAA) polypeptide, wherein the heterologous nucleic acid is operatively linked to a liver specific promoter.wherein the recombinant AAV vector comprises a capsid protein of the AAV3b or AAV8 serotype.35. The recombinant AAV vector of claim 34 , wherein the polypeptide further comprises a secretory signal peptide located at the N-terminal of the GAA polypeptide.36. The recombinant AAV vector of ...

SYNTHETIC PROMOTERS AND USES THEREOF

The present invention relates to the treatment and/or prevention of a retinal disease by using a polynucleotide promoter wherein the polynucleotide or a variant thereof consists of the sequence 1- (canceled)2- The polynucleotide promoter or a variant thereof according to claim 4 , wherein said promoter has a promoter activity at least 40% higher or at least 25% lower than the wild-type promoter of SEQ ID No. 1.7- The polynucleotide or a variant thereof according to wherein said polynucleotide or a variant thereof shows a promoter activity in retina cells claim 4 , photoreceptors claim 4 , or rods.8- A vector comprising the polynucleotide or a variant thereof according to .9- A vector comprising the polynucleotide or a variant thereof according to .10- A vector comprising a first expression cassette comprising the polynucleotide or a variant thereof according to and a first transgene under the control of said polynucleotide.11- The vector of claim 10 , wherein said first transgene encodes for a transcriptional repressor.12- The vector according to wherein said transcriptional repressor is selected from the group consisting of: an antisense oligonucleotide claim 11 , a siRNA claim 11 , a shRNA or a miRNA claim 11 , targeting a RHO transcript; an artificial transcription factor (ATF) comprising a DNA Binding domain coupled to one or more effector domains claim 11 , targeting a sequence of the hRHO promoter; an isolated DNA Binding domain (DNA binding domain or DBD) claim 11 , and targeting a sequence of the hRHO promoter.13- The vector according to claim 10 , comprising a further expression cassette claim 10 , said further expression cassette comprises a further promoter and a further transgene under control of said further promoter claim 10 , optionally wherein said further promoter is a polynucleotide which is the same or it is different from the polynucleotide of the first expression cassette.14- The vector according to wherein the further transgene is a nucleotide ...

Therapeutic retroviral vectors for gene therapy

Retroviral gene therapy vectors that are optimized for erythroid specific expression and treatment of hemoglobinopathic conditions are disclosed.

Brain-Specific Enhancers for Cell-Based Therapy

Herein are described a set of novel specific human enhancers for specific forebrain cell types used to study and select for human neural progenitor cells. This approach enables the ability to generate interneurons from human ES, iPS and iN cells, making them available for human transplantation and for molecular/cellular analyses. These approaches are also directly applicable to generating other neuronal cell types, such as cortical and striatal projection neurons, which have implications for many human diseases.

Treatment methods using adenovirus

The invention provides methods of reducing or decreasing a size of a tumor or eliminating a tumor or inhibiting, decreasing, or reducing neo-vascularization or angiogenesis in a tumor in a patient by administering an adenovirus comprising a nucleic acid construct comprising a FAS-chimera gene operably linked to an endothelial cell-specific promoter. Also provided is a homogeneous population of an adenovirus comprising a FAS-chimera gene operably linked to an endothelial cell-specific promoter and its uses thereof.

LIVER-SPECIFIC NUCLEIC ACID REGULATORY ELEMENTS AND METHODS AND USE THEREOF

Described are nucleic acid regulatory elements that are able to enhance liver-specific expression of genes, methods employing these regulatory elements and uses of these elements. Expression cassettes and vectors containing these nucleic acid regulatory elements are also disclosed. These are particularly useful for applications using gene therapy. 120.-. (canceled)21. A nucleic acid expression cassette comprising: wherein the nucleic acid regulatory element comprises SEQ ID NO:14 or a polynucleotide having at least 95% identity to SEQ ID NO:14, and', 'wherein the nucleic acid regulatory element is operably linked to a promoter and a heterologous transgene., 'a nucleic acid regulatory element of 200 nucleotides or less that enhances liver-specific gene expression,'}22. The nucleic acid expression cassette of claim 21 , further comprising:at least one additional nucleic acid regulatory element that enhances liver-specific gene expression comprising a polynucleotide having at least 95% identity to a sequence selected from the group consisting of: SEQ ID NO:1, SEQ ID NO:2, SEQ ID NO:3, SEQ ID NO:4, SEQ ID NO:5, SEQ ID NO:6, SEQ ID NO:7, SEQ ID NO:8, SEQ ID NO:9, SEQ ID NO:10, SEQ ID NO:11, SEQ ID NO:12, SEQ ID NO:13, and SEQ ID NO:14, wherein said second nucleic acid regulatory element is 600 nucleotides or less.23. The nucleic acid expression cassette of claim 22 , wherein the total length of the combined nucleic acid regulatory elements does not exceed 700 nucleotides.24. The nucleic acid expression cassette of claim 22 , wherein the cassette comprises at least two identical nucleic acid regulatory elements.25. The nucleic acid expression cassette of claim 22 , wherein the cassette comprises repeats of polynucleotides having at least 95% identity to SEQ ID NO:14.26. The nucleic acid expression cassette of claim 25 , wherein the cassette comprises three repeats of polynucleotides having at least 95% identity to SEQ ID NO:14.27. The nucleic acid expression cassette of ...

GENE AUGMENTATION THERAPIES FOR INHERITED RETINAL DEGENERATION CAUSED BY MUTATIONS IN THE PRPF31 GENE

The present invention relates to methods and compositions for gene therapy of retinitis pigmentosa related to mutations in pre-mRNA processing factor 31 (PRPF31). 1. A method of treating retinitis pigmentosa caused by mutations in PRPF31 in a human subject , the method comprising delivering to the eye of the subject a therapeutically effective amount of an Adeno-associated virus type 2 (AAV2) vector comprising a sequence encoding human PRPF31 , operably linked to a promoter that drives expression in retinal pigment epithelial (RPE) cells.2. The method of wherein the promoter is a CAG claim 1 , CASI claim 1 , RPE65 or VMD2 promotor.3. The method of claim 2 , wherein the PRPF31 sequence is codon optimized.4. The method of claim 1 , wherein the vector is delivered via sub-retinal injection.5. A method of increasing expression of PRPF31 in the eye of a human subject claim 1 , the method comprising delivering to the eye of the subject a therapeutically effective amount of an Adeno-associated virus type 2 (AAV2) vector comprising a sequence encoding human PRPF31 claim 1 , operably linked to a promoter that drives expression in retinal pigment epithelial (RPE) cells.6. The method of claim 5 , wherein the promoter is a CAG claim 5 , CASI claim 5 , RPE65 or VMD2 promotor.7. The method of claim 5 , wherein the PRPF31 sequence is codon optimized.8. The method of claim 5 , wherein the vector is delivered via sub-retinal injection.9. An Adeno-associated virus type 2 (AAV2) vector comprising a sequence encoding human PRPF31 claim 5 , operably linked to a promotor that drives expression in retinal pigment epithelial (RPE) cells.10. The vector of claim 9 , wherein the promotor is a CAG claim 9 , CASI claim 9 , RPE65 or VMD2 promotor.11. The vector of claim 9 , wherein the PRPF31 sequence is codon optimized.12. A pharmaceutical composition comprising the vector of claim 9 , formulated for delivery via sub-retinal injection.13. The vector of claim 9 , for use in treating retinitis ...

ADENOVIRAL TARGETING, COMPOSITIONS AND METHODS THEREFOR

Polypeptides are disclosed comprising, in N-terminal-to-C-terminal order: an N-terminal segment of Ad5 fiber tail sequence; at least 2 pseudorepeats of an Ad5 fiber shaft domain sequence; a portion of a third Ad5 fiber shaft domain sequence; a carboxy-terminal segment of a 14 fibritin bacteriophage trimerization domain sequence; a linker sequence; and a camelid single chain antibody sequence. A camelid single chain antibody sequence can be against a human carcinoembryonic antigen. Also disclosed are nucleic acids encoding these polypeptides, and adenovirus vectors comprising the polypeptides. Methods are disclosed for treating a neoplastic disease. These methods can comprise administering an adenovirus vector comprising a disclosed polypeptide. Also disclosed are methods of targeting a vector to CEA-expressing cells. These methods comprise administering an adenovirus vector comprising a disclosed polypeptide. Methods can further comprise subjecting a subject to ionizing radiation in an amount effective for inducing CEA overexpression. 1. A polypeptide comprising , in N-terminal-to-C-terminal order;an N-terminal segment of Ad5 fiber tail sequence;at least 2 pseudorepeats of an Ad5 fiber shaft domain sequence;a portion of a third Ad5 fiber shaft domain sequence;a carboxy-terminal segment of a T4 fibritin bacteriophage trimerization domain sequence;a linker sequence; anda camelid single chain antibody sequence.2. A polypeptide in accordance with claim 1 , wherein the carboxy-terminal segment of the T4 fibritin bacteriophage trimerization domain sequence comprises an α-helicaJ domain and a foldon domain.3. A polypeptide in accordance with claim 1 , wherein the N-terminal segment of Ad5 fiber tail sequence is a sequence having at least 70% sequence identity with SEQ ID NO.1.4. A polypeptide in accordance with claim 1 , wherein the at least 2 pseudorepeats of an Ad5 fiber shaft domain sequence is a sequence having at least 70% sequence identity with SEQ ID NO:2.5. A ...

Engineered Cellular Pathways for Programmed Autoregulation of Differentiation

The present invention provides compositions and methods for programming mammalian cells to perform desired functions. In particular, the present invention provides compositions and methods for programming stem cells to differentiate into a desired cell type. A quorum sensing systems that regulates the expression of cell fate regulators is introduced into mammalian host cells, such as stem cells. The quorum sensing systems generally comprises vectors that express the components of a bacterial quorum sensing pathway, including proteins which catalyze the synthesis of an autoinducer and a gene encoding a regulatory partner of the autoinducer, and vectors in which genes encoding cell fate regulators are operably linked to a promoter induced by the autoinducer/regulatory partner complex. The system can also comprise vectors in which genes encoding additional cell fate regulators are operably linked to a promoter that is induced by a factor synthesized in response to a first stage of differentiation, so that a second stage of differentiation is triggered.

USE AND PRODUCTION OF CHD8+/- TRANSGENIC ANIMALS WITH BEHAVIORAL PHENOTYPES CHARACTERISTIC OF AUTISM SPECTRUM DISORDER

The invention involves inducing a plurality e.g., 3-50 or more mutations (e.g., any whole number between 3 and 50 or more of mutations, with it noted that in some embodiments there can be up to 16 different RNA(s), e.g., sgRNAs each having its own a promoter, in a vector, such an AAV vector or a lentiviral vector and that when each sgRNA does not have its own promoter, there can be twice to thrice that amount of different RNA(s), e.g., sgRNAs, e.g., 32 or even 48 different guides delivered by one vector) in transgenic Cas9 eukaryotes to model a neuronal disease or disorder. The invention comprehends testing putative treatments with such models, e.g., testing putative chemical compounds that may be pharmaceutically relevant for treatment or gene therapy that may be relevant for treatment, or combinations thereof. The invention allows for the study of genetic diseases and putative treatments to better understand and alleviate a genetic disease or a condition, e.g., autism, autism-spectrum disease or disorder, obsessive compulsive disorder, or psychiatric disorders. 176-. (canceled)77. A method for modeling a neuronal/nervous system related disease or disorder , comprising introducing multiple mutations ex vivo in a tissue , organ or a cell line comprising Cas9-expressing eukaryotic cell(s) or cell(s) that are able to be induced to express or that conditionally express Cas9 , or in vivo in a transgenic non-human mammal having cells that express or that are able to be induced to express or that conditionally express Cas9 , comprising delivering to cell(s) of the tissue , organ , cell or mammal a vector which expresses a plurality of guides to guide the Cas9 to a plurality of neuronal cell-specific target loci and optionally deliver donor templates and a plurality of neuronal cell-specific mutations or precise sequence substitutions.78. The method of claim 77 , wherein the specific mutations or precise sequence substitutions are or have been correlated to the neuronal/ ...

Codon optimized rep1 genes and uses thereof

The present disclosure provides codon optimized nucleotide sequences encoding human REP1, vectors, and host cells comprising codon optimized REP1 sequences, and methods of treating retinal disorders such as choroideremia comprising administering to the subject a codon optimized sequence encoding human REP1.

LIVER-SPECIFIC NUCLEIC ACID REGULATORY ELEMENTS AND METHODS AND USE THEREOF

Described are nucleic acid regulatory elements that are able to enhance liver-specific expression of genes, methods employing these regulatory elements and uses of these elements. Expression cassettes and vectors containing these nucleic acid regulatory elements are also disclosed. These are particularly useful for applications using gene therapy. 119.-. (canceled)20. A nucleic acid expression cassette comprising:a nucleic acid regulatory element of 100 nucleotides or less that enhances liver-specific gene expression,wherein the nucleic acid regulatory element comprises SEQ ID NO:7, or a polynucleotide having at least 95% identity to SEQ ID NO:7, andwherein the nucleic acid regulatory element is operably linked to a promoter and a heterologous transgene.21. The nucleic acid expression cassette of claim 20 , wherein the nucleic acid regulatory element does not form part of a larger regulatory region.22. A nucleic acid expression cassette comprising: a first polynucleotide of 100 nucleotides or less comprising SEQ ID NO:7 or a polynucleotide having at least 95% identity to SEQ ID NO:7; and', 'a second nucleic acid regulatory element of 90 nucleotides or less comprising SEQ ID NO:3 or a polynucleotide having at least 95% identity to SEQ ID NO:3., 'at least two nucleic acid regulatory elements that enhance liver-specific gene expression, each of the at least two nucleic acid regulatory elements comprising23. The nucleic acid expression cassette of claim 22 , wherein the at least two nucleic acid regulatory elements do not form part of a larger regulatory region.24. The nucleic acid expression cassette of claim 22 , wherein the length of the total of the at least two regulatory elements does not exceed 700 nucleotides.25. The nucleic acid expression cassette of claim 22 , wherein the at least two nucleic acid regulatory elements are identical.26. The nucleic acid expression cassette of claim 22 , wherein the at least two nucleic acid regulatory elements comprise repeats ...

Bicistronic AAV Vector for RNA Interference in ALS

The present invention relates to a bicistronic expression vector for silencing a gene specifically in astrocytes and neurons, comprising two expression cassettes comprising a first and a second silencer sequence, respectively, wherein the expression of said first silencer sequence within astrocytes is regulated by an astrocyte-specific promoter and the expression of said second silencer sequence within neurons is regulated by a neuron-specific promoter. In a preferred embodiment, said first and second silencer sequences are SOD1 silencer sequences. Pharmaceutical composition comprising said bicistronic vector and the use of the same in the treatment of motoneuron diseases are further described. 1. A method of treating a motoneuron disease comprising:administering to a subject in need thereof a bicistronic expression vector for silencing a gene specifically in astrocytes and neurons, a first expression cassettes comprising a first silencer sequence, and', 'a second expression cassette comprising a second silencer sequence,, 'wherein the bicistronic expression vector comprises'}wherein the expression of said first silencer sequence within astrocytes is regulated by an astrocyte-specific promoter and the expression of said second silencer sequence within neurons is regulated by a neuron-specific promoter.2. The method according to claim 1 , wherein said bicistronic expression vector comprises two expression cassettes claim 1 , whereina first cassette comprises “astrocyte specific promoter-posttranscriptional regulatory element-the first silencer sequence-polyA tail,” anda second cassette comprises “neuron specific promoter-posttranscriptional regulatory element-the second silencer sequence-polyA tail” for silencing a gene in astrocytes and neurons.3. The method according to claim 1 , wherein said first and second silencer sequences are both superoxide dismutase 1 (SOD1) silencer sequences.4. The method according to claim 3 , wherein said SOD1 silencer sequences are ...

Transdermal Optogenetic Peripheral Nerve Stimulation

A nerve in a mammal is optogenetically transduced, wherein the nerve is susceptible to stimulus by selective application of transdermal light, and a light source is applied to dermis of the mammal at or proximate to the optogenetically transduced nerve, to thereby stimulate the nerve. A wearable device for optogenetic motor control and sensation restoration of a mammal includes a wearable support, a power source at the wearable support, a controller at the wearable support and in electrical communication with a power source, and a transdermal light source coupled to the controller. 1. A wearable device for optogenetic motor control and restoring sensation in a mammal , comprising:a) a wearable support;b) a power source at the wearable support;c) a controller at the wearable support and in electrical communication with the power source; andd) a transdermal light source coupled to the controller, the controller driving the light source to direct light from the wearable support and directed toward the mammal while wearing the support.2. The wearable device of claim 1 , wherein the wearable support is a strap.3. The wearable device of claim 2 , wherein the strap is a member selected from the group consisting of a wrist strap claim 2 , a knee strap claim 2 , a necklace claim 2 , a headband claim 2 , ankle strap claim 2 , leg strap claim 2 , stomach strap claim 2 , and an arm strap.4. The wearable device of claim 1 , wherein the wearable support is an adhesive patch.5. The wearable device of claim 1 , wherein the transdermal light source includes at least one member selected from the group consisting of a light emitting diode (LED) claim 1 , diode-pumped solid state (DPSS) laser claim 1 , diode laser claim 1 , solid-state laser claim 1 , vertical-cavity surface emitting laser (VCSEL) claim 1 , and edge emitting laser diode (EELD).6. The wearable device of claim 1 , wherein the light source is of a type that emits a wavelength in a range of between about 300 nm and about ...

PRIMATE RETINAL PIGMENT EPITHELIUM CELL-SPECIFIC PROMOTER

The present invention to a method for expressing an exogenous gene specifically in cells of the retinal pigment epithelium N of a primate, this method comprising the step of delivering an isolated nucleic acid molecule comprising, or consisting of, the nucleic acid sequence of SEQ ID NO:1, or consisting of a nucleic acid sequence of at least 400 bp having at least 80% overall identity to the sequence of SEQ ID NO:1, to cells of the retinal pigment epithelium of the primate, wherein this isolated nucleic acid molecule specifically leads to the expression of an exogenous gene in cells of the retinal pigment epithelium of primates when a nucleic acid sequence coding for the exogenous gene is operatively linked to this isolated nucleic acid molecule. 125-. (canceled)26. A method for expressing an exogenous gene in a retinal pigment epithelial cell of a primate , the method comprising delivering to the retinal pigment epithelial cell of the primate an expression cassette comprising a nucleic acid sequence of at least 400 bp and having at least 95% overall identity to the nucleic acid sequence of SEQ ID NO:1 , operatively linked to a nucleic acid sequence coding for the exogenous gene ,wherein the nucleic acid sequence of at least 400 bp and having at least 95% overall identity to the nucleic acid sequence of SEQ ID NO:1 is effective to drive expression of the exogenous gene in the retinal pigment epithelial cell of the primate.27. The method of claim 26 , wherein the nucleic acid sequence of at least 400 bp and having at least 95% overall identity to the nucleic acid sequence of SEQ ID NO:1 comprises the nucleic acid sequence of SEQ ID NO:1.28. The method of claim 26 , wherein the expression cassette further comprises a minimal promoter sequence.29. The method of claim 28 , wherein the minimal promoter sequence comprises the nucleic acid sequence of SEQ ID NO:2.30. The method of claim 26 , wherein the exogenous gene encodes a channelrhodopsin or halorhodopsin.31. A ...

Methods to modulate protein translation efficiency

The present disclosure relates to compositions and methods to modulate the level of expression of a protein in a deliberate manner (i.e., tunable regulation of expression) with only a minimal change to the genetic sequence of the gene of interest. The present disclosure therefore also provides compositions and methods to predictably alter protein abundance.

COMPOSITIONS AND METHODS FOR TREATING DISEASES

The present invention provides compositions and methods of use pertaining to rAAV-mediated delivery of therapeutically effective molecules for treatment of diseases such as Pompe disease. These compositions in combination with various routes and methods of administration result in targeted expression of therapeutic molecules in specific organs, tissues and cells. 1. A method of improving impaired neuromuscular junction integrity , comprising administering , to a subject with impaired neuromuscular junction integrity , an effective amount of a composition comprising a rAAV 2/9 vector , wherein the rAAV2/9 vector comprises a heterologous nucleic acid molecule operably linked to a promoter , and the therapeutic composition is administered to the subject via intramuscular , intrathoracic , intraspinal , intrathecal , or intravenous injection.2. The method according to claim 1 , wherein the heterologous nucleic acid molecule encodes acid α-glucosidase (GAA).3. The method according to claim 1 , wherein the promoter is a cytomegalovirus (CMV) promoter or a desmin (DES) promoter.4. The method according to claim 1 , wherein the promoter is a desmin (DES) promoter.5. The method according to claim 1 , wherein the subject has impaired neuromuscular caused by a neuromuscular disease.6. The method according to claim 1 , wherein the subject has impaired neuromuscular caused by a disease selected from the group consisting of Pompe disease claim 1 , amyotrophic lateral sclerosis claim 1 , spinal muscular atrophy claim 1 , multiple sclerosis claim 1 , glycogen storage disease type 1a claim 1 , limb Girdle muscular dystrophy claim 1 , Barth syndrome claim 1 , and myasthenia gravis.7. The method according to claim 1 , wherein the subject is a human.8. A method of treating a neuromuscular disease claim 1 , comprising administering claim 1 , to a subject in need of such treatment claim 1 , an effective amount of a composition comprising a rAAV 2/9 vector claim 1 , wherein the rAAV2/9 ...

NOVEL METHOD FOR LABELLING CANCERIZATION-CAUSING CELL IN STEM CELLS, AND THERAPY METHOD

The invention provides a method to search for a best promoter for reliably targeting a cancerization-causing cell in human ES/iPS cells. Discovering a gene which can kill or remove a cancerization-causing cell contained in human ES/iPS cells with high efficiency is also provided. A method is provided for removing an undifferentiated cell, which remains after the differentiation of human ES/iPS cells into desired cells, with high efficiency and comprehensively. A viral vector is provided which includes a nucleotide sequence and a recombination cassette. The nucleotide sequence contains a target gene and a killing gene that are linked to each other through a sequence that enables the simultaneous expression of the two genes by one promoter. The recombination cassette contains a promoter region which is so linked as to enable the expression of the labeling gene and the killing gene. The viral vector may contain a promoter specific to an undifferentiated cell. 1. A viral vector comprising a nucleic acid sequence in which a marker gene and a toxic gene are bound via a sequence capable of allowing a promoter to cause the two genes to be simultaneously expressed and a recombination cassette having a promoter region , wherein the promoter region is operably linked to the marker gene and the toxic gene.2. The viral vector according to claim 1 , wherein the toxic gene is a suicide gene.3. The viral vector according to claim 2 , wherein the suicide gene is a drug-dependent suicide gene.4. The viral vector according to claim 3 , wherein the drug-dependent suicide gene is HSV-tk claim 3 , human-tmpk claim 3 , a cytosine deaminase gene claim 3 , herpes virus thymidine kinase claim 3 , or caspase.5. The viral vector according to claim 1 , wherein the marker gene is a fluorescent protein.6. The viral vector according to claim 5 , wherein the fluorescent protein is a red fluorescent protein.7. The viral vector according to claim 5 , wherein the fluorescent protein is mKate2.8. The ...

SELECTIVE GENE THERAPY EXPRESSION SYSTEM

The present invention relates to an expression system for systemic administration comprising a sequence encoding a protein, said expression system allowing: 1. An expression system for systemic administration comprising a sequence encoding a protein having a therapeutic effect on neuromuscular disorders and whose expression is toxic in at least one tissue , said expression system allowing:the expression at a therapeutically acceptable level of the protein in the target tissues including skeletal muscles and/or the peripheral nervous tissue; andthe expression at toxically acceptable level of the protein in tissues other than the target tissues, especially in the heart.2. The expression system according to claim 1 , wherein it comprises at least one sequence allowing:the prevention of the expression or the reduction in the level of expression of the protein in tissues other than the target tissues, preferably those in which the expression of the protein is toxic; and/orthe maintenance of the expression or the increase in the level of expression of the protein in the target tissues.3. The expression system according to claim 1 , wherein the protein is myotubularin claim 1 , preferably with the sequence SEQ ID NO: 1 claim 1 , 2 or 3.4. The expression system according to claim 1 , wherein the protein is calpain 3 claim 1 , preferably with the sequence SEQ ID NO: 7.5. The expression system according to claim 1 , wherein it comprises a target sequence of an miRNA expressed in tissues other than the target tissues claim 1 , preferably those in which the expression of the protein is toxic.6. The expression system according to wherein it comprises at least one target sequence of the miR208a claim 5 , preferably of sequence SEQ ID NO: 10.7. The expression system according to claim 1 , wherein it comprises a promoter sequence presenting low or no promoter activity in tissues other than the target tissues claim 1 , preferably those in which the expression of the protein is toxic ...

TREATMENT METHODS USING ADENOVIRUS

The invention provides methods of reducing or decreasing a size of a tumor or eliminating a tumor by inhibiting, decreasing, or reducing neo-vascularization or angiogenesis in a tumor in a patient by administering an adenovirus comprising a nucleic acid construct comprising a FAS-chimera gene operably linked to an endothelial cell-specific promoter. Also provided is a homogeneous population of an adenovirus comprising a FAS-chimera gene operably linked to an endothelial cell-specific promoter and its uses thereof. 1107.-. (canceled)108. A promoter comprising the sequence set forth in nucleotides 1 to 987 of SEQ ID NO: 18.109. A vector comprising the promoter of claim 108 , wherein the vector comprises the sequence set forth in nucleotides 1 to 35207 of SEQ ID NO: 19.110. A pharmaceutical composition comprising the vector of and a pharmaceutically acceptable carrier.111. A method of producing the vector of claim 109 , the method comprising transducing a host cell with the vector and expressing the vector in the host cell.112. The method of claim 111 , wherein the host cell comprises an adenovirus E1 region.113. The method of claim 112 , wherein the host cell is a mammalian cell.114. An isolated mammalian cell transfected with the vector of .115. A method of inhibiting claim 109 , reducing claim 109 , or decreasing a size of a tumor in a subject in need thereof claim 109 , the method comprising administering to the subject an effective amount of the vector of claim 109 , wherein expression of the vector inhibits claim 109 , reduces claim 109 , or decreases the size of the tumor.116. The method of claim 115 , further comprising administering an effective amount of one or more chemotherapeutic agents.117. The method of claim 116 , wherein the one or more chemotherapeutic agents are selected from altretamine claim 116 , raltritrexed claim 116 , topotecan claim 116 , paclitaxel claim 116 , docetaxel claim 116 , cisplatin claim 116 , carboplatin claim 116 , oxaliplatin ...

ADENO-ASSOCIATED VIRUS (AAV) CLADES, SEQUENCES, VECTORS CONTAINING SAME, AND USES THEREFOR

Sequences of novel adeno-associated virus capsids and vectors and host cells containing these sequences are provided. Also described are methods of using such host cells and vectors in production of rAAV particles. AAV-mediated delivery of therapeutic and immunogenic genes using the vectors of the invention is also provided. 1. A recombinant nucleic acid molecule:(a) encoding an AAV9 vp1 capsid protein having a sequence comprising amino acids 1 to 736 of SEQ ID NO: 123; or(b) comprising nucleotides 1 to 2208 of SEQ ID NO: 3, or a nucleotide sequence at least 99% identical to nucleotides 1 to 2208 of SEQ ID NO: 3,wherein the recombinant nucleic acid molecule does not contain an AAV inverted terminal repeat.2. The recombinant nucleic acid molecule according to claim 1 , which further comprises a functional rep gene.3. The recombinant nucleic acid molecule according to claim 1 , wherein said recombinant nucleic acid molecule comprises a sequence selected from:vp1, nt 1 to 2208;vp2, nt 412 to 2208; orvp3, nt 607 to 2208 of SEQ ID NO: 3.4. The recombinant nucleic acid molecule according to claim 1 , wherein said recombinant nucleic acid molecule is a plasmid.5. A host cell transfected with the recombinant nucleic acid molecule according to .6. The host cell according to claim 5 , which further comprises a functional rep gene claim 5 , a minigene comprising AAV inverted terminal repeats (ITRs) and a transgene claim 5 , and sufficient helper functions to permit packaging of the minigene into the AAV capsid.7. The host cell according to claim 6 , which further comprises a nucleic acid sequence(a) encoding an AAV9 vp2 capsid protein having a sequence comprising amino acids 138 to 736 of SEQ ID NO: 123; or(b) comprising nucleotides 412 to 2208 of SEQ ID NO: 3, or a nucleotide sequence at least 99% identical to nucleotides 412 to 2208 of SEQ ID NO: 3.8. The host cell according to claim 6 , which further comprises a nucleic acid sequence(a) encoding an AAV9 vp3 capsid protein ...

SYNP161, A PROMOTER FOR THE SPECIFIC EXPRESSION OF GENES IN ROD PHOTORECEPTORS

The present invention provides an isolated nucleic acid molecule comprising, or consisting of, the nucleic acid sequence of SEQ ID NO:1 or a nucleic acid sequence of at least 150 bp having at least 80% identity to said sequence of SEQ ID NO:1, wherein said isolated nucleic acid molecule specifically leads to the expression in rod photoreceptors of a gene when operatively linked to a nucleic acid sequence coding for said gene. 1. An isolated nucleic acid molecule comprising , or consisting of , the nucleic acid sequence of SEQ ID NO:1 , or consisting of a nucleic acid sequence of at least 150 bp having at least 80% identity to said sequence of SEQ ID NO:1 , wherein said isolated nucleic acid molecule leads to the specific expression of a gene in rod photoreceptors when a nucleic acid sequence coding for said gene is operatively linked to said isolated nucleic acid molecule.2. The isolated nucleic acid molecule of claim 1 , further comprising a minimal promoter.3. An isolated nucleic acid molecule comprising a sequence that hybridizes under stringent conditions to an isolated nucleic acid molecule according to .4. An expression cassette comprising claim 1 , as an element promoting gene expression in specific cells claim 1 , an isolated nucleic acid according to claim 1 , wherein said isolated nucleic acid is operatively linked to at least a nucleic acid sequence encoding for a gene to be expressed specifically in rod photoreceptors.5. A vector comprising the expression cassette of .6. The vector of claim 5 , wherein said vector is a viral vector.7. (canceled)8. A method of a expressing gene in rod photoreceptors comprising the steps of transfecting an isolated cell claim 4 , a cell line or a cell population with an expression cassette according to claim 4 , wherein the gene to be expressed will be specifically expressed by the isolated cell claim 4 , the cell line or the cell population if said cell is claim 4 , or said cells comprise claim 4 , rod photoreceptors.9. ...

LIVER-SPECIFIC VIRAL PROMOTERS AND METHODS OF USING THE SAME

The present invention relates to promoters that function specifically or preferentially in the liver. These promoters are capable of enhancing liver-specific expression of genes. The invention also relates to expression constructs, vectors and cells comprising such liver-specific promoters, and to methods of their use. The present invention future relates to adeno-associated virus (AAV) gene therapy vectors comprising the liver-specific promoters, therapeutic agents comprising the liver-specific promoters, and methods using the same. 1. A synthetic polynucleotide , comprising:{'claim-text': ['(a) HNF1/HNF3 (SEQ ID NO:1),', '(b) HNF3/HNF3 (SEQ ID NO:2),', '(c) c/EBP/HNF4 (SEQ ID NO:3),', '(d) HS_CRM2/HNF3 (SEQ ID NO:4), and', '(e) HS_CRM8 (SEQ ID NO:6) or a variant thereof, or'], '#text': '(I) at least three promoter-derived nucleic acids selected from the group consisting of:'}{'claim-text': ['(f) Motif_44 (SEQ ID NO:12),', '(g) NRF2F1 (SEQ ID NO:14),', '(h) HNF1A (SEQ ID NO:15), and,', '(i) IA2 (SEQ ID NO:16); or'], '#text': '(II) at least three promoter-derived nucleic acids selected from the group consisting of:'}(III) a sequence selected from the group consisting of SEQ ID Nos: 36, 37, 38, 39, and 40; or at least 90% identity with such a sequence.2. The synthetic polynucleotide according to claim 1 , comprising at least SEQ ID NO:3 and SEQ ID NO:6 or a variant thereof.3. The synthetic polynucleotide according to claim 1 , wherein the variant of the HS_CRM8 sequence is selected from the group consisting of SEQ ID NO:5 claim 1 , SEQ ID NO:87 claim 1 , SEQ ID NO:88 claim 1 , SEQ ID NO:89 claim 1 , SEQ ID NO:90 claim 1 , SEQ ID NO:91 claim 1 , SEQ ID NO:92 claim 1 , SEQ ID NO:93 claim 1 , SEQ ID NO:94 claim 1 , SEQ ID NO:95 claim 1 , SEQ ID NO:96 claim 1 , SEQ ID NO:97 claim 1 , SEQ ID NO:98 claim 1 , SEQ ID NO:99 claim 1 , and SEQ ID NO: 100.4. The synthetic polynucleotide according to claim 1 , comprising at least four of the promoter-derived nucleic acids.5. The ...

AAV/XBP1S-HA VIRUS, GENE THERAPY METHOD AND USE THEREOF IN THE OPTIMISATION AND IMPROVEMENT OF LEARNING, MEMORY AND COGNITIVE CAPACITIES

This invention presents a sequence of the virus AAV/XBP1s-HA, method and its use in the improvement of cognitive functions, of memory and of learning, as presented in the in vivo studies in FIG. right panel. 1. A method of treating a disease related to the memory , cognitive or learning capacity in a mammal subject in need thereof , said method comprising administering a therapeutically efficient amount of a viral vector that induces neuronal overexpression of XBP1 in the brain , wherein said viral vector comprises an expression cassette with a nucleotide sequence encoding the neuronal transcription factor XBP1.2. The method of claim 1 , wherein said neuronal overexpression of XBP1 improves the performance of the memory claim 1 , cognitive or learning capacities in said mammal.3. The method of claim 1 , wherein said XBP1 is XBP1s.4. The method of claim 1 , wherein the viral vector is of the adeno-associated type (AAV).5. The method of claim 1 , wherein the viral vector is administered into the hippocampus.6. The method of claim 1 , wherein said viral vector is an AAV vector comprising a recombinant genome with the expression cassette including a transcriptional regulating region specific of hippocampal tissue operatively linked to the nucleotide sequence encoding XBP1 s.7. The method of claim 1 , wherein the virus is an AAV vector comprising an insert of a nucleotide encoding XBP1 s as contained in the plasmid deposited at the ATCC under deposit number PTA-121708.8. The method according to claim 1 , wherein the viral vector is an AAV vector selected from AAV6 claim 1 , AAV7 claim 1 , AAV8 and AAV9.9. The method according to claim 6 , wherein the serotype of the AAV is AAV6.10. The method according to claim 1 , wherein the nucleotide sequence encoding XBP1 is operably linked to the regulating region of the specific transcription of neuronal tissue selected from Pgkl claim 1 , Cam 2 and Thy 1.11. The method according to claim 1 , wherein the expression cassette ...

EXPRESSION SYSTEMS

A gene expression system is provided. The system comprises at least one coding sequence to be expressed in an organism, and at least one promoter operably linked thereto. It further comprises at least one splice control sequence which, in cooperation with a spliceosome, mediates alternative splicing of RNA transcripts of the coding sequence. The mediation of alternative splicing is in a sex-specific, stage-specific, germline-specific and tissue-specific manner. 1. A gene expression system comprising at least one coding sequence to be expressed in an organism, at least one promoter operably linked thereto, and at least one splice control sequence which, in cooperation with a spliceosome, mediates alternative splicing of RNA transcripts of the coding sequence, the mediation being selected from at least one of the group consisting of: sex-specific, stage-specific, germline-specific and tissue-specific mediation. This application is a continuation of U.S. application Ser. No. 11/352,177, filed Feb. 10, 2006, now pending, which is a continuation-in-part of U.S. application Ser. No. 10/566,448, filed Apr. 18, 2006, now pending, which is the national stage entry of International Application No. PCT/GB2004/003263, filed Jul. 28, 2004, which claims the priority from GB 0317656.7, filed Jul. 28, 2003. All applications are hereby incorporated by reference in their entireties for all purposes.The content of the following submission on ASCII text file is incorporated herein by reference in its entirety: a computer readable form (CRF) of the Sequence Listing (file name: 750402000401SeqList.txt, date recorded: Aug. 26, 2015, size: 434,690 bytes).The present invention relates to a gene expression system, in combination with splice control sequences, said control sequences providing a mechanism for alternative splicing.Alternative splicing is also known as pre-mRNA splicing and involves the removal of one or more introns and ligation of the flanking exons. This reaction is catalyzed ...

REGENERATING FUNCTIONAL NEURONS FOR TREATMENT OF NEURAL INJURY CAUSED BY DISRUPTION OF BLOOD FLOW

Methods of treating the effects of disruption of normal blood flow in the CNS in an individual subject in need thereof are provided according to aspects of the present invention which include administering a therapeutically effective dose of exogenous NeuroD1 to an area where normal blood flow has been disrupted. Compositions are provided including 1) a recombinant adeno-associated adenovirus expression vector comprising a glial cell specific promoter operably linked to a nucleic acid encoding a site-specific recombinase and 2) a recombinant adeno-associated adenovirus expression vector comprising a ubiquitous promoter operably linked to a nucleic acid encoding NeuroD1, wherein the nucleic acid encoding NeuroD1 is inverted and flanked by two sets of site-specific recombinase recognition sites such that action of the recombinase irreversibly inverts the nucleic acid encoding NeuroD1 such that NeuroD1 is expressed in a mammalian cell. 1. A method of treating the effects of disruption of normal blood flow in the CNS in an individual subject in need thereof , comprising:administering a therapeutically effective dose of exogenous NeuroD1 to an area where normal blood flow has been disrupted.2. The method of claim 1 , wherein administering exogenous NeuroD1 comprises delivering an expression vector comprising a nucleic acid encoding NeuroD1 to the area.3. The method of or claim 1 , wherein administering exogenous NeuroD1 comprises delivering a recombinant viral expression vector comprising a nucleic acid encoding NeuroD1 to the area.4. The method of claim 1 , wherein expressing exogenous NeuroD1 comprises delivering a recombinant adeno-associated virus expression vector comprising a nucleic acid encoding NeuroD1 to the area.516-. (canceled)17. The method of claim 1 , further comprising assessing the effectiveness of the treatment in the subject.18. The method of claim 17 , wherein assessing the effectiveness of the treatment in the subject comprises an assay selected from ...

RAAV VECTOR COMPOSITIONS, METHODS FOR TARGETING VASCULAR ENDOTHELIAL CELLS AND USE IN TREATMENT OF TYPE I DIABETES

Disclosed are capsid-modified rAAV expression vectors, as well as infectious virions, compositions, and pharmaceutical formulations that include them. Also disclosed are methods of preparing and using novel capsid-protein-mutated rAAV vector constructs in a variety of diagnostic and therapeutic applications including, inter alia, as delivery agents for diagnosis, treatment, or amelioration of one or more diseases, disorders, or dysfunctions of the mammalian vascular system, and complications from Type I diabetes. Also disclosed are methods for systemic and tissue-localized delivery of therapeutic rAAV-based gene expression cassettes to vascular endothelial cells, tissues, and organs, as well as use of the disclosed compositions in the manufacture of medicaments for a variety of in vitro and/or in vivo applications including the treatment of vasculitis, and complications arising from Type I diabetes, such as macular edema, nephropathy, diabetic retinopathy, and the like. 1. A recombinant adeno-associated viral (rAAV) expression system comprising:a) a polynucleotide that encodes a modified capsid protein, wherein the modified capsid protein comprises five or more non-native amino acid substitutions at positions corresponding to five or more distinct surface-exposed amino acid residues in the wild-type AAV2 capsid protein, and further wherein the transduction efficiency of a virion comprising the modified capsid protein is higher than that of a virion comprising a corresponding, unmodified wild-type capsid protein; andb) an expression cassette packaged within the virion, that comprises an isolated polynucleotide comprising a nucleic acid segment that encodes or that expresses a diagnostic or a therapeutic molecule in a mammal transformed with the expression system, wherein the nucleic acid segment is operably linked to a promoter or a control region that expresses the nucleic acid segment in one or more vascular endothelial cells of the mammal to produce the diagnostic ...

CHIMERIC GENE CONSTRUCTS FOR GENERATION OF FLUORESCENT TRANSGENIC ORNAMENTAL FISH

Four zebrafish gene promoters, which are skin specific, muscle specific, skeletal muscle specific and ubiquitously expressed respectively, were isolated and ligated to the 5′ end of the EGFP gene. When the resulting chimeric gene constructs were introduced into zebrafish, the transgenic zebrafish emit green fluorescence under a blue light or ultraviolet light according to the specificity of the promoters used. Thus, new varieties of ornamental fish of different fluorescence patterns, e.g., skin fluorescence, muscle fluorescence, skeletal muscle-specific and/or ubiquitous fluorescence, are developed. 141-. (canceled)42. A transgenic fish comprising a chimeric gene comprising a promoter operably linked to an exogenous gene , wherein said promoter comprises a nucleic acid sequence that is identical to the sequence of SEQ ID NO: 7 , SEQ ID NO: 8 , SEQ ID NO: 9 , or SEQ ID NO: 22 , wherein the transgenic fish contains said promoter in germ cells and/or in somatic cells and is capable of breeding to produce viable and fertile transgenic progeny that express the exogenous gene.43. The transgenic fish of claim 42 , wherein the variant polypeptide or variant polynucleotides comprises at least 75% sequence identity to the naturally occurring sequences or fragments.44. The transgenic fish of claim 42 , wherein the variant polypeptide or variant polynucleotides comprises at least 85% sequence identity to the naturally occurring sequences or fragments.45. The transgenic fish of claim 42 , wherein the variant polypeptide or variant polynucleotides comprises at least 95% sequence identity to the naturally occurring sequences or fragments.46. The transgenic fish of claim 42 , wherein the variant polypeptide or variant polynucleotides comprises at least 98% sequence identity to the naturally occurring sequences or fragments.47. The transgenic fish of claim 42 , further comprising a fluorescent protein gene under control of said promoter.48. The transgenic fish of claim 47 , wherein ...

RECOMBINANT CONSTRUCTS AND TRANSGENIC FLUORESCENT ORNAMENTAL FISH THEREFROM

The present invention relates to the method and use of reef coral fluorescent proteins in making transgenic red, green and yellow fluorescent zebrafish. Preferably, such fluorescent zebrafish are fertile and used to establish a population of transgenic zebrafish and to provide to the ornamental fish industry for the purpose of marketing. Thus, new varieties of ornamental fish of different fluorescence colors from a novel source are developed. 171-. (canceled)72. A transgenic fluorescent fish comprising in its genome: (a) a first transgene encoding a first fluorescent protein , wherein the first transgene is under the control of a tissue specific promoter , and (b) a second transgene encoding a second fluorescent protein , wherein the second transgene is under the control of a tissue specific promoter; wherein the first and second transgene encode different fluorescent proteins , wherein the fluorescent fish visibly express a fluorescent color different from the color encoded by the first and second transgene.73. The transgenic fluorescent fish of claim 72 , wherein the transgenic fluorescent fish exhibit a strong visible fluorescence over 75% to 100% of the body of said fish claim 72 , optionally excluding fins and eyes.74. The transgenic fluorescent fish of claim 72 , wherein said first and second transgenes are both chromosomally integrated at the same claim 72 , single locus.75. The transgenic fluorescent fish of claim 72 , further comprising a third transgene encoding a third fluorescent protein claim 72 , wherein said third transgene is under the control of a ubiquitous promoter.76. The transgenic fluorescent fish of claim 72 , wherein the first transgene encodes for the color red.77. The transgenic fluorescent fish of claim 72 , wherein the second transgene encodes for the color yellow.78. The transgenic fluorescent fish of claim 72 , wherein the first transgene encodes for the color red and the second transgene encodes for the color yellow.79. A transgenic ...

REPLICATION-DEFECTIVE ARENAVIRUS VECTORS

The invention relates to an infectious arenavirus particle that is engineered to contain a genome with the ability to amplify and express its genetic information in infected cells but unable to produce further infectious progeny particles in normal, not genetically engineered cells. One or more of the four arenavirus open reading frames glycoprotein (GP), nucleoprotein (NP), matrix protein Z and RNA-dependent RNA polymerase L are removed or mutated to prevent replication in normal cells but still allowing gene expression in arenavirus vector-infected cells, and foreign genes coding for an antigen or other protein of interest or nucleic acids modulating host gene expression are expressed under control of the arenavirus promoters, internal ribosome entry sites or under control of regulatory elements that can be read by the viral RNA-dependent RNA polymerase, cellular RNA polymerase I, RNA polymerase II or RNA polymerase III. The modified arenaviruses are useful as vaccines and therapeutic agents for a variety of diseases. 1. An infectious arenavirus particle engineered to contain a genome with the ability to amplify and express its genetic information in infected cells but unable to produce further infectious progeny particles in normal , not genetically engineered cells.2. The arenavirus particle according to comprising additional nucleic acids coding for a protein or peptide of interest.3. The arenavirus particle according to comprising additional nucleic acids modulating host gene expression.4. The arenavirus particle according to comprising a modified genome claim 2 , whereini) one or more of the four arenavirus open reading frames glycoprotein (GP), nucleoprotein (NP), matrix protein Z and RNA-dependent RNA polymerase L are removed or mutated to prevent replication in normal cells but still allowing gene expression in arenavirus vector-infected cells;ii) foreign ribonucleic acids coding for one or more proteins of interest are expressed under control of one or ...

Engineered Cellular Pathways for Programmed Autoregulation of Differentiation

The present invention provides compositions and methods for programming mammalian cells to perform desired functions. In particular, the present invention provides compositions and methods for programming stem cells to differentiate into a desired cell type. A quorum sensing systems that regulates the expression of cell fate regulators is introduced into mammalian host cells, such as stem cells. The quorum sensing systems generally comprises vectors that express the components of a bacterial quorum sensing pathway, including proteins which catalyze the synthesis of an autoinducer and a gene encoding a regulatory partner of the autoinducer, and vectors in which genes encoding cell fate regulators are operably linked to a promoter induced by the autoinducer/regulatory partner complex. The system can also comprise vectors in which genes encoding additional cell fate regulators are operably linked to a promoter that is induced by a factor synthesized in response to a first stage of differentiation, so that a second stage of differentiation is triggered. 177-. (canceled)78. A composition comprising one or more mammalian vectors that comprise:a) a first nucleic acid sequence capable of producing a first cell fate regulator protein that is capable of inducing differentiation of a first cell type into a second cell type that expresses a protein marker, andb) a second nucleic acid sequence capable of producing a second cell fate regulator protein that is operably linked to a cell type specific promoter of said second cell type, and that is capable of inducing differentiation of said second cell type into a third cell type79. The composition of claim 78 , wherein said one or more vectors further comprisesc) a third nucleic acid sequence capable of producing a third cell fate regulator protein that is operably linked to a cell type specific promoter of said second cell type, and that is capable of inducing differentiation of said second cell type into said third cell type.80. ...

METHODS AND COMPOSITIONS RELATING TO RESTRICTED EXPRESSION LENTIVIRAL VECTORS AND THEIR APPLICATIONS

The present invention provides HIV-derived lentivectors which are safe, highly efficient, and very potent for expressing transgenes for human gene therapy, especially, in human hematopoietic progenitor cells as well as in all other blood cell derivatives. The lentiviral vectors comprise promoters active to promote expression specific to cell types or tissues. Further, promoters are providing that are amenable to control by activators, enhancers, or repressors. These vectors are in a self-inactivating configuration for biosaftey. Additional promoters are also described. The vectors can also comprise additional transcription enhancing elements such as the wood chuck hepatitis virus post-transcriptional regulatory element, without any decrease in the specificity or control exerted by the promoters. These vectors therefore provide useful tools for genetic treatments such as inherited and acquired lympho-hematological disorders, gene-therapies for cancers especially the hematological cancers, as well as for the study of hematopoiesis via lentivector-mediated modification of human HSCs. 168.-. (canceled)69. A lymphocyte transduced in vitro with a lentivirus comprising a transgene positioned under the control of a promoter that is active to support detectable transcription of the transgene in the lymphocyte , and capable of promoting expression of the transgene in the lymphocyte at a signal-to-noise ratio of between about 10 and about 200.70. The lymphocyte of claim 69 , wherein the lymphocyte is a T lymphocyte.71. The lymphocyte of claim 69 , wherein the lentivirus comprises a central polypurine tract (cPPT) positioned upstream of the transgene.72. The lymphocyte of claim 71 , wherein the central polypurine tract (cPPT) comprises the nucleotide sequence of SEQ ID NO: 1.73. The lymphocyte of claim 71 , wherein the lentivirus comprises multiple unique cloning sites positioned adjacent to the central polypurine tract (cPPT).74. The lymphocyte of claim 73 , wherein the ...

Optimized Liver-Specific Expression Systems for FVIII and FIX

The present invention relates to nucleic acid expression cassettes and vectors containing liver-specific regulatory elements and codon-optimized factor IX or factor VIII transgenes, methods employing these expression cassettes and vectors and uses thereof. The present invention is particularly useful for applications using liver-directed gene therapy, in particular for the treatment of hemophilia A and B. 1. A nucleic acid expression cassette comprising a triple repeat of a liver-specific nucleic acid regulatory element comprising the nucleic acid fragment defined by SEQ ID NO:5 or a sequence having at least 95% identity to said sequence; and a nucleic acid regulatory element comprising the nucleic acid fragment defined by SEQ ID NO: 12 or a sequence having at least 95% identity to said sequence; operably linked to a liver-specific promoter and a transgene.2. The nucleic acid expression cassette according to claim 1 , wherein the liver-specific promoter is selected from the group comprising: the minimal TTR promotor (TTRm) claim 1 , the AAT promoter claim 1 , the albumin (ALB) promotor or minimal promoter claim 1 , the apolipoprotein A1 (APOA1) promoter or minimal promoter claim 1 , the complement factor B (CFB) promoter claim 1 , the ketohexokinase (KHK) promoter claim 1 , the hemopexin (HPX) promoter or minimal promoter claim 1 , the nicotinamide N-methyltransferase (NNMT) promoter or minimal promoter claim 1 , the (liver) carboxylesterase 1 (CES1) promoter or minimal promoter claim 1 , the protein C (PROC) promoter or minimal promoter claim 1 , the apolipoprotein C3 (APOC3) promoter or minimal promoter claim 1 , the mannan-binding lectin serine protease 2 (MASP2) promoter or minimal promoter claim 1 , the hepcidin antimicrobial peptide (HAMP) promoter or minimal promoter claim 1 , or the serpin peptidase inhibitor claim 1 , clade C (antithrombin) claim 1 , member 1 (SERPINC1) promoter or minimal promoter.3. The nucleic acid expression cassette according to claim ...

Compositions and methods for the treatment of degenerative ocular diseases

The present invention provides compositions, e.g., pharmaceutical compositions, which include a recombinant adeno-associated viral (AAV) expression construct, AAV vectors, AAV particles, and methods of treating a subject having a degenerative ocular disorder, e.g., retinitis pigmentosa.

Methods of treating solid or lymphatic tumors by combination therapy

The present invention provides methods for treating an individual having solid or lymphatic tumor comprising locally administering to the site of the tumor an oncolytic virus, and systemically administering an immunomodulator (including a combination of immunomodulators). The methods may further comprise local administration to the site of the tumor a second immunomodulator (including a combination of immunomodulators). Also provided are compositions and kits for the cancer therapy methods.

Non-invasive in vivo imaging and methods for treating diabetes

The present invention provides novel drug discovery platforms and methods for treating diabetes. 1. A method for treating a subject with diabetes , comprising transplanting into an eye of a subject with diabetes an amount effective of insulin-producing cells to promote insulin production in the subject.2. The method of claim 1 , wherein the insulin-producing cells are present in an isolated pancreatic islet.3. The method of claim 1 , wherein the insulin-producing cells comprise isolated pancreatic β cells.4. The method of claim 1 , wherein the transplantation into the eye involves transplantation onto the iris of the eye.5. The method of claim 1 , wherein the treating comprises restoring normoglycemia.6. The method of claim 2 , wherein the transplantation into the eye involves transplantation onto the iris of the eye.7. The method of claim 3 , wherein the transplantation into the eye involves transplantation onto the iris of the eye.8. The method of claim 2 , wherein the treating comprises restoring normoglycemia.9. The method of claim 3 , wherein the treating comprises restoring normoglycemia.10. The method of claim 4 , wherein the treating comprises restoring normoglycemia.11. The method of claim 6 , wherein the treating comprises restoring normoglycemia.12. The method of claim 7 , wherein the treating comprises restoring normoglycemia. This application is a continuation of U.S. patent application Ser. No. 15/265,212 filed Sep. 14, 2016, which is a continuation of U.S. patent application Ser. No. 14/515,000 filed Oct. 15, 2014, now U.S. Pat. No. 9,463,205 issued Oct. 11, 2016, which is a divisional of U.S. application Ser. No. 12/199,473 filed Aug. 27, 2008, which claims priority to U.S. Provisional Patent Application Ser. No. 60/969,437 filed Aug. 31, 2007, 61/042,482 filed Apr. 4, 2008, and 60/989,038 filed Nov. 19, 2007, all incorporated by reference herein in their entirety.Fundamental understanding of cellular processes in health and disease has been gained ...

Lineage reporter synthetic chromosomes and methods of use

The field of the invention encompasses synthetic chromosome compositions and methods that allow single cell spatiotemporal analysis in response to differentiation cues and labeling of transplanted cells to monitor the fate and function of such cells in the patient recipient.

Materials and Methods for the Treatment of Pathological Neovascularization in the Eye

The subject invention provides materials and methods useful in safely and effectively preventing pathological proliferation of blood vessels. The prevention of the over-proliferation of blood vessels according to the subject invention is particularly advantageous for treatment of certain ocular conditions including age-related macular degeneration (AMD), retinopathy of prematurity (ROP) and diabetic retinopathy. In preferred embodiments, the subject invention provides materials and methods for effective treatment of pathological ocular neovascularization using gene therapy. In a specific embodiment the materials and methods of the subject invention can be used to treat AMD.