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Небесная энциклопедия

Космические корабли и станции, автоматические КА и методы их проектирования, бортовые комплексы управления, системы и средства жизнеобеспечения, особенности технологии производства ракетно-космических систем

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Мониторинг СМИ

Мониторинг СМИ и социальных сетей. Сканирование интернета, новостных сайтов, специализированных контентных площадок на базе мессенджеров. Гибкие настройки фильтров и первоначальных источников.

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Форма поиска

Поддерживает ввод нескольких поисковых фраз (по одной на строку). При поиске обеспечивает поддержку морфологии русского и английского языка
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Применить Всего найдено 8405. Отображено 100.
19-04-2012 дата публикации

Human artificial chromosome (hac) vector

Номер: US20120093785A1
Автор: Kazuma Tomizuka, Minoru Kakeda, Mitsuo Oshimura, Motonobu Katoh, Yoshimi Kuroiwa
Принадлежит: Kirin Brewery Co Ltd

The present invention relates to a human artificial chromosome (HAC) vector and a method for producing the same. The present invention further relates to a method for introducing foreign DNA using a human artificial chromosome vector and a method for producing a cell which expresses foreign DNA. Furthermore, the present invention relates to a method for producing a protein.

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Номер записи: 1
25-10-2012 дата публикации

Reverse genetics systems

Номер: US20120270321A1
Автор: Michael Franti, Peter Mason, Philip Dormitzer
Принадлежит: NOVARTIS AG

The invention provides various reverse genetics systems for producing segmented RNA viruses, wherein the systems do not require bacteria for propagation of all of their expression constructs.

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Номер записи: 2
20-12-2012 дата публикации

Adam6 Mice

Номер: US20120322108A1
Автор: Andrew J. Murphy, Lynn Macdonald, Sean Stevens
Принадлежит: Regeneron Pharmaceuticals Inc

Mice are provided that comprise a reduction or deletion of ADAM6 activity from an endogenous ADAM6 locus, or that lack an endogenous locus encoding a mouse ADAM6 protein, wherein the mice comprise a sequence encoding an ADAM6 or ortholog or homolog or fragment thereof that is functional in a male mouse. In one embodiment, the sequence is an ectopic ADAM6 sequence or a sequence that confers upon a male mouse the ability to generate offspring by mating. Mice and cells with genetically modified immunoglobulin heavy chain loci that comprise an ectopic nucleotide sequence encoding a mouse ADAM6 or functional fragment or homolog or ortholog thereof are also provided.

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Номер записи: 3
04-04-2013 дата публикации

Method for the identification of t cell epitopes

Номер: US20130085260A1
Автор: Jean-Daniel Doucet, Rejean Lapointe
Принадлежит: Centre Hospitalier de lUniversite de Montreal CHUM

A novel method to identify relevant T-cell epitopes recognized by CD8 + or CD4 − T lymphocytes is described. The method is based on the use of mRNA fragments synthesized from cDNA encoding portions of a polypeptide of interest. mRNA fragments are introduced into antigen-presenting cells to deduce an epitope's localization in a polypeptide of interest, such as a protein antigen.

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Номер записи: 4
20-06-2013 дата публикации

Optimized promoter sequence

Номер: US20130158246A1
Автор: Gary Kobinger, Heinz Feldmann, Kaylie Tran
Принадлежит: Canada Minister of National Health and Welfare

A modified CAG promoter which is capable of driving high levels of expression of sequences of interest inserted downstream therefrom is herein described.

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Номер записи: 5
18-07-2013 дата публикации

Computationally optimized broadly reactive antigens for influenza

Номер: US20130183342A1
Автор: Brendan M. Giles, Ted M. Ross
Принадлежит: Individual

Described herein is the development of a computationally optimized influenza HA protein that elicits broadly reactive immune response to all H5N1 influenza virus isolates. The optimized HA protein was developed through a series of HA protein alignments, and subsequent generation of consensus sequences, for clade 2 H5N1 influenza virus isolates. The final consensus HA amino acid sequence was reverse translated and optimized for expression in mammalian cells. It is disclosed herein that influenza virus-like particles containing the optimized HA protein are an effective vaccine against H5N1 influenza virus infection in animals.

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Номер записи: 6
01-08-2013 дата публикации

Poxvirus Expression System

Номер: US20130195912A1
Автор: Matthew Guy Cottingham
Принадлежит: Oxford University Innovation Ltd

There is provided a method for inserting a nucleic acid sequence that encodes a foreign peptide into a poxvirus genome, said method comprising: identifying in the poxvirus genome a poxvirus open reading frame wherein said open reading frame is characterised by an initial ATG start codon and wherein expression of said open reading frame is driven by an operably-linked poxvirus promoter located upstream of the open reading frame and wherein expression of said open reading frame provides a peptide that is non-essential to viability of the poxvirus; and inserting the nucleic acid sequence that encodes the foreign peptide at a position downstream of the poxvirus promoter; wherein following said insertion, (i) the nucleic acid that encodes the foreign peptide is operably-linked to the poxvirus promoter and expression of said nucleic acid is driven by said poxvirus promoter; and (ii) translation of the foreign peptide is initiated at an ATG start codon located at the same position as the ATG start codon of the poxvirus open reading frame. Also provided are a poxvirus vector and corresponding uses of the poxvirus vector in medicine.

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Номер записи: 7
19-09-2013 дата публикации

Non-Human Animals Expressing pH-Sensitive Immunoglobulin Sequences

Номер: US20130247236A1
Автор: Andrew J. Murphy, Joel H. Martin, John Mcwhirter, Lynn Macdonald
Принадлежит: Regeneron Pharmaceuticals Inc

Genetically modified non-human animals are provided that express an immunoglobulin variable domain that comprises at least one histidine, wherein the at least one histidine is encoded by a substitution of a non-histidine codon in the germline of the animal with a hisidine codon, or the insertion of a histidine codon in a germline immunoglobulin nucleic acid sequence. Immunoglobulin genes comprising histidines in one or more CDRs, in an N-terminal region, and or in a loop 4 region are also provided. Immunoglobulin variable domains comprising one or more histidines (e.g., histidine clusters) substituted for non-antigen-binding non-histidine residues. Non-human animals that are progeny of animals comprising modified heavy chain variable loci (V, D, J segments), modified light chain variable loci (V, J segments), and rearranged germline light chain genes (VJ sequences) are also provided. Non-human animals that make immunoglobulin domains that bind antigens in a pH-sensitive manner are provided.

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Номер записи: 8
17-10-2013 дата публикации

Production of Transgenic Avians Using Improved Retroviral Vectors

Номер: US20130276153A1
Автор: Alex J. Harvey, Jeffrey C. Rapp
Принадлежит: Alex J. Harvey, Jeffrey C. Rapp

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.

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Номер записи: 9
21-11-2013 дата публикации

Promoter-regulated differentiation-dependent self-deleting cassette

Номер: US20130312128A1
Автор: David Frendewey, David M. Valenzuela, Guochun Gong, Ka-Man Venus Lai
Принадлежит: Regeneron Pharmaceuticals Inc

Targeting constructs and methods of using them are provided for differentiation-dependent modification of nucleic acid sequences in cells and in non-human animals. Targeting constructs comprising a promoter operably linked to a recombinase are provided, wherein the promoter drives transcription of the recombinase in an differentiated cell but not an undifferentiated cell. Promoters include Blimp1, Prm1, Gata6, Gata4, Igf2, Lhx2, Lhx5, and Pax3. Targeting constructs with a cassette flanked on both sides by recombinase sites can be removed using a recombinase gene operably linked to a 3′-UTR that comprises a recognition site for an miRNA that is transcribed in undifferentiated cells but not in differentiated cells. The constructs may be included in targeting vectors, and can be used to automatically modify or excise a selection cassette from an ES cell, a non-human embryo, or a non-human animal.

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Номер записи: 10
05-12-2013 дата публикации

Restricted immunoglobulin heavy chain mice

Номер: US20130323791A1
Автор: Andrew J. Murphy, Cagan Gurer, John Mcwhirter, Karolina A. Hosiawa, Lynn Macdonald
Принадлежит: Regeneron Pharmaceuticals Inc

Mice having a restricted immunoglobulin heavy chain locus are provided, wherein the locus is characterized by a single polymorphic human V H gene segment, a plurality of human D H gene segments and a plurality of J H gene segments. Methods for making antibody sequences that bind an antigen (e.g., a viral antigen) are provided, comprising immunizing a mouse with an antigen of interest, wherein the mouse comprises a single human V H gene segment, a plurality of human D H gene segments and a plurality of J H gene segments, at the endogenous immunoglobulin heavy chain locus.

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Номер записи: 11
12-12-2013 дата публикации

Humanized Non-Human Animals with Restricted Immunoglobulin Heavy Chain Loci

Номер: US20130333057A1
Автор: Andrew J. Murphy, John Mcwhirter, Lynn Macdonald, Naxin Tu, Sean Stevens
Принадлежит: Regeneron Pharmaceuticals Inc

Mice, embryos, cells, and tissues having a restricted immunoglobulin heavy chain locus and an ectopic sequence encoding one or more ADAM6 proteins are provided. In various embodiments, mice are described that have humanized endogenous immunoglobulin heavy chain loci and are capable of expressing an ADAM6 protein or ortholog or homolog or functional fragment thereof that is functional in a male mouse. Mice, embryos, cells, and tissues having an immunoglobulin heavy chain locus characterized by a single human V H gene segment, a plurality of human D H gene segments and a plurality of human J H gene segments and capable expressing an ADAM6 protein or ortholog or homolog or functional fragment thereof are also provided.

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Номер записи: 12
27-02-2014 дата публикации

Polynucleotide and polypeptide sequence and methods thereof

Номер: US20140057318A1
Автор: Chandrasekhar Bhaskaran Nair, Kirubakaran Naveen Kumar, Madhuri Baliga, MULAKKAPURATH NARAYANAN Manoj, Pillarisetti Venkata Subbarao, Venkata Ramachandra Rao Vasamsetty
Принадлежит: Bigtec Pvt Ltd

The present disclosure relates to a field of recombinant DNA therapeutics. It involves the bio-informatics design, synthesis of artificial gene for human insulin precursor including leader peptide coding sequence, cloning in an expression vector and expression in an organism, preferably Pichia pastoris. The present disclosure also relates to methods of downstream processing for obtaining protein precursor molecules and subsequent conversion of precursor molecules to functional proteins.

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Номер записи: 13
07-01-2021 дата публикации

Optimised Coding Sequence and Promoter

Номер: US20210000978A1
Автор: Davidoff Andrew, Gray John, McVey John, Nathwani Amit, Thrasher Adrian, Tuddenham Edward, Ward Natalie
Принадлежит:

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 ...

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Номер записи: 14
02-01-2020 дата публикации

METHOD FOR THE TREATMENT OF MALIGNANCIES

Номер: US20200000938A1
Автор: DAUD ADIL, Pierce Robert H.
Принадлежит:

The present invention provides for the intratumoral delivery of at least one immunostimulatory cytokine in combination with at least one checkpoint inhibitor. In particular, it provides delivery of a plasmid encoding the immunostimulatory cytokine using intratumoral electroporation. The checkpoint inhibitor may be administered systemically or encoded on a plasmid and delivered using intratumoral electroporation. The checkpoint inhibitor may be delivered contemporaneously with or after treatment with the immunomodulatory cytokine. 1. A method of treating a subject having a cancerous tumor , the method comprising:a) injecting the cancerous tumor with an effective dose of at least one plasmid coding for at least one immunostimulatory cytokine;b) administering electroporation therapy to the tumor; andc) administering an effective dose of a checkpoint inhibitor to the subject.2. The method of claim 1 , wherein the electroporation therapy comprises the administration of at least one voltage pulse over a duration of about 100 microseconds to about 1 millisecond.3. The method of claim 1 , wherein the at least one voltage pulse delivered to the tumor has a field strength of about 20 V/cm to about 1500 V/cm.4. The method of claim 1 , wherein the checkpoint inhibitor is administered systemically.5. The method of wherein the checkpoint inhibitor is encoded on a plasmid and delivered to the cancerous tumor by electroporation therapy.6. The method of claim 1 , wherein the checkpoint inhibitor is encoded on the plasmid encoding the immunostimulatory cytokine and delivered to the cancerous tumor by electroporation therapy.7. The method of claim 1 , wherein the checkpoint inhibitor is an antagonist of at least one checkpoint target of Table 1.8. The method of claim 7 , wherein the checkpoint inhibitor is selected from the group consisting of: nivolumab (ONO-4538/BMS-936558 claim 7 , MDX1106 claim 7 , OPDIVO) claim 7 , pembrolizumab (MK-3475 claim 7 , KEYTRUDA) claim 7 , pidilizumab ...

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Номер записи: 15
02-01-2020 дата публикации

TREATMENT OF RETINITIS PIGMENTOSA

Номер: US20200002392A1
Автор: Fischer Dominik, MacLaren Robert
Принадлежит:

A polynucleotide comprising a nucleotide sequence encoding the retinitis pigmentosa GTPase regulator ORF15 isoform (RPGR), wherein the RPGR-encoding nucleotide sequence has been codon optimised to increase fidelity of replication of the sequence. 1. A composition comprising a nucleic acid comprising(a) a sequence encoding a rhodopsin kinase promoter, and{'sup': 'ORF15', '(b) a sequence encoding a retinitis pigmentosa GTPase regulator ORF15 isoform (RPGR) and comprising a nucleotide sequence that has at least 80% identity to the nucleic acid sequence of SEQ ID NO: 3.'}2. The composition of claim 1 , wherein the sequence encoding the promoter has at least 80% identity to the nucleic acid sequence of SEQ ID NO: 7.3. The composition of claim 2 , wherein the sequence encoding the promoter has at least 80% identity to the nucleic acid sequence of SEQ ID NO: 7.4. The composition of claim 1 , wherein the sequence encoding the promoter comprises the nucleic acid sequence of SEQ ID NO: 7.5. The composition of claim 2 , wherein the sequence encoding the promoter comprises the nucleic acid sequence of SEQ ID NO: 7.6. The composition of claim 1 , wherein the sequence encoding the promoter consists of the nucleic acid sequence of SEQ ID NO: 7.7. The composition of claim 2 , wherein the sequence encoding the promoter consists of the nucleic acid sequence of SEQ ID NO: 7.8. The composition of claim 1 , wherein the sequence encoding the RPGRcomprises a nucleotide sequence encoding an amino acid sequence that has at least 80% identity to the amino acid sequence of SEQ ID NO: 1.9. The composition of claim 2 , wherein the sequence encoding the RPGRcomprises a nucleotide sequence encoding an amino acid sequence that has at least 80% identity to the amino acid sequence of SEQ ID NO: 1.10. The composition of claim 3 , wherein the sequence encoding the RPGRcomprises a nucleotide sequence encoding an amino acid sequence that has at least 80% identity to the amino acid sequence of SEQ ID NO: ...

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Номер записи: 16
04-01-2018 дата публикации

Collagen 7 and related methods

Номер: US20180002401A1
Автор: Malini Viswanathan, Mark De Souza
Принадлежит: Phoenix Tissue Repair Inc

Disclosed are methods of making collagen 7, or functional fragments thereof, as well as collagen 7, and functional fragments thereof produced by such methods, nucleic acids encoding collagen 7, and functional fragments thereof, as well as vectors and host cells comprising such nucleic acids.

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Номер записи: 17
04-01-2018 дата публикации

Engineered CRISPR-Cas9 nucleases with Altered PAM Specificity

Номер: US20180002681A9
Автор: Joung J. Keith, Kleinstiver Benjamin
Принадлежит:

Engineered CRISPR-Cas9 nucleases with altered and improved PAM specificities and their use in genomic engineering, epigenomic engineering, and genome targeting. 1Staphylococcus aureus. An isolated Cas9 (SaCas9) protein with mutations at one or more of the following positions: E782 , N968 , and/or R1015.2. The isolated protein of claim 1 , comprising a sequence that is at least 80% identical to the amino acid sequence of SEQ ID NO:2.3. The isolated protein of claim 1 , comprising one or more of the following mutations: R1015Q claim 1 , R1015H claim 1 , E782K claim 1 , N968K claim 1 , E735K claim 1 , K929R claim 1 , A1021T claim 1 , K1044N claim 1 , E782K/N968K/R1015H (KKH variant); E782K/K929R/R1015H (KRH variant); or E782K/K929R/N968K/R1015H (KRKH variant).4. The isolated protein of claim 1 , further comprising one or more mutations that decrease nuclease activity selected from the group consisting of mutations at one or more of D10 claim 1 , D556 claim 1 , H557 claim 1 , and/or N580.5. The isolated protein of claim 4 , wherein the mutations are one or more of D10A claim 4 , D556A claim 4 , H557A claim 4 , or N580A.6. The isolated protein of claim 4 , wherein the mutations are:(i) D10A/H557A, or(ii) D10A/D556A/H557A/N580A.7. A fusion protein comprising the isolated protein of claim 1 , fused to a heterologous functional domain claim 1 , with an optional intervening linker claim 1 , wherein the linker does not interfere with activity of the fusion protein.8. The fusion protein of claim 7 , wherein the heterologous functional domain is a transcriptional activation domain.9. The fusion protein of claim 8 , wherein the transcriptional activation domain is from VP64 or NF-κK p65.10. The fusion protein of claim 7 , wherein the heterologous functional domain is a transcriptional silencer or transcriptional repression domain.11. The fusion protein of claim 10 , wherein the transcriptional repression domain is a Krueppel-associated box (KRAB) domain claim 10 , ERF repressor ...

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Номер записи: 18
03-01-2019 дата публикации

Plant promoter for transgene expression

Номер: US20190002910A1
Автор: Andrew ASBERRY, Andrew J. Bowling, Daren Hemingway, Emily Etchison, Heather Pence, Pierluigi Barone, Sandeep Kumar
Принадлежит: DOW AGROSCIENCES LLC

This disclosure concerns compositions and methods for promoting transcription of a nucleotide sequence in a plant or plant cell, employing a promoter from a Panicum virgatum (Pavir.Cb02009) egg cell gene. Some embodiments relate to a promoter from a Panicum virgatum (Pavir.Cb02009) egg cell gene that functions in plants to promote transcription of operably linked nucleotide sequences. Other embodiments relate to a 3′ UTR from a Panicum virgatum (Pavir.Cb02009) egg cell gene that functions in plants to promote transcription of operably linked nucleotide sequences.

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Номер записи: 19
03-01-2019 дата публикации

METHODS AND COMPOSITIONS FOR TRANSPOSITION USING MINIMAL SEGMENTS OF THE EUKARYOTIC TRANSFORMATION VECTOR PIGGYBAC

Номер: US20190002914A1
Автор: Fraser Malcolm, LI Xu
Принадлежит:

Isolated nucleic acid molecules that include a minimally-sized, functional (minimally-functional) piggyBac transposon can incorporate (i) a 5′ internal domain (ID) comprising a nucleotide fragment that is substantially homologous to a native piggyBac transposon sequence; (ii) a 5′ terminal repeat domain (TRD) comprising a 5′ terminal repeat (TR) sequence, a 5′ spacer sequence, and a 5′ internal repeat (IR) sequence; (ii) a sequence of interest; (iv) a 3′ TRD comprising a 3′ IR sequence, a 3′ spacer sequence, and a 3′ TR sequence; and (iv) a 3′ ID comprising a nucleotide fragment that is substantially homologous to the native piggyBac transposon sequence. The 5′ TRD and the 3′ TRD can be optionally linked by a sequence comprising a multiple cloning site. 1. An isolated nucleic acid molecule comprising a minimally-sized , functional (minimally-functional) piggyBac transposon , the minimally-functional piggyBac transposon comprising the following genetic elements in a 5′ to 3′ direction:a 5′ internal domain (ID) comprising a nucleotide fragment that is substantially homologous to a native piggyBac transposon sequence;a 5′ terminal repeat domain (TRD) comprising a 5′ terminal repeat (TR) sequence, a 5′ spacer sequence, and a 5′ internal repeat (IR) sequence;a sequence of interest;a 3′ TRD comprising a 3′ IR sequence, a 3′ spacer sequence, and a 3′ TR sequence; anda 3′ ID comprising a nucleotide fragment that is substantially homologous to the native piggyBac transposon sequence.2. The isolated nucleic acid molecule of claim 1 , wherein the 5′ TRD and the 3′ TRD are linked by a sequence comprising a multiple cloning site.3. The isolated nucleic acid molecule of claim 1 , wherein the nucleotide fragments of the 5′ ID and the 3′ ID are each at least 66-bp in length.4. The isolated nucleic acid molecule of claim 1 , wherein the nucleotide fragment of the 5′ ID corresponds to a sequence selected from the group consisting of: SEQ ID NO: 192 claim 1 , SEQ ID NO: 198 claim 1 , ...

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Номер записи: 20
20-01-2022 дата публикации

FABRY DISEASE GENE THERAPY

Номер: US20220016263A1
Автор: Nathwani Amit, Raj Deepak
Принадлежит:

There is described a nucleic acid molecule comprising a nucleotide sequence encoding for a functional α-galactosidase A protein wherein the nucleotide sequence has at least 85% identity to the sequence of SEQ ID NO. 1. Also described is a vector, host cell or transgenic animal comprising the nucleic acid molecule; and a pharmaceutical composition comprising the nucleic acid molecule or the vector. Further, the use of the nucleic acid molecule in a method of treating Fabry disease is described. 117-. (canceled)18. A method of administering a nucleic acid molecule to a subject , comprising administering to the subject the nucleic acid molecule comprising a nucleotide sequence encoding a polypeptide having α-galactosidase A activity , wherein the nucleotide sequence has at least 91% identity to the nucleotide sequence of SEQ ID NO: 1.191. The method of claim , wherein the subject suffers from Fabry disease.201. The method of claim , wherein the nucleotide sequence has at least 92% identity to the nucleotide sequence of SEQ ID NO: 1.211. The method of claim , wherein the nucleotide sequence has at least 93% identity to the nucleotide sequence of SEQ ID NO: 1.221. The method of claim , wherein the nucleotide sequence has at least 94% identity to the nucleotide sequence of SEQ ID NO: 1.231. The method of claim , wherein the nucleotide sequence has at least 95% identity to the nucleotide sequence of SEQ ID NO: 1.241. The method of claim , wherein the nucleotide sequence has at least 96% identity to the nucleotide sequence of SEQ ID NO: 1.251. The method of claim , wherein the nucleotide sequence has at least 97% identity to the nucleotide sequence of SEQ ID NO: 1.261. The method of claim , wherein the nucleotide sequence has at least 98% identity to the nucleotide sequence of SEQ ID NO: 1.271. The method of claim , wherein the nucleotide sequence has at least 99% identity to the nucleotide sequence of SEQ ID NO: 1.281. The method of claim , wherein the nucleotide sequence ...

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Номер записи: 21
20-01-2022 дата публикации

EXPOSED COLLAGEN-TARGETED FUSION CYTOKINE FOR IMMUNE MODULATION IN INVASIVE CANCERS AND LESIONS OF INFECTIONS

Номер: US20220017586A1
Автор: Gordon Erlinda M., Hall Frederick L.
Принадлежит:

Provided herein are new compositions and methods to target pharmaceutical agents to pathological areas by utilizing fusion polypeptides. These fusion polypeptides contain two or more domains: (i) aptamer sequences that bind to exposed collagenous (XC-) proteins present in pathological areas, including cancerous and viral lesions, (ii) immunomodulators, such as cytokines, and optionally (iii) at least one linker joining the two domains or at the terminus of the polypeptide. In some cases, the linker is a rigid linker, e.g., a rigid helical linker. Also provided herein are methods of treating cancer and/or infectious diseases using the new fusion polypeptides. 184-. (canceled)85. A method of treating an infection in a subject comprising administering to a subject in need of such treatment a fusion polypeptide comprising an amino acid sequence selected from the group consisting of:APARSPSPSTQPWEHVNAIQEARRLLNLSRDTAAEMNETVEVISEMFDLQEPTCLQTRLE LYKQGLRGSLTKLKGPLTMMASHYKQHCPPTPETSCATQIITFESFKENLKDFLLVIPFDC WEPVQEGAEAAAKEAAAKAGARRGVRVAWREPGRMELNMPHGQE (SEQ ID NO: 17);APARSPSPSTQPWEHVNAIQEARRLLNLSRDTAAEMNETVEVISEMFDLQEPTCLQTRLE LYKQGLRGSLTKLKGPLTMMASHYKQHCPPTPETSCATQIITFESFKENLKDFLLVIPFDC WEPVQEGSAGSAAGSGARRGVRVAWREPGRMELNMPHGQE (SEQ ID NO: 18);METDTLLLWVLLLWVPGSTGREEFEEEFEHREEFEFEENLYFQGAPARSPSPSTQPWEHVNAIQE ARRLLNLSRDTAAEMNETVEVISEMFDLQEPTCLQTRLELYKQGLRGSLTKLKGPLTMM ASHYKQHCPPTPETSCATQIITFESFKENLKDFLLVIPFDCWEPVQEGAEAAAKEAAAKA GARRGVRVAWREPGRMELNMPHGQE (SEQ ID NO: 19);1VIETDTLLLWVLLLWVPGSTGREEFEEEFEHREEFEFEENLYFQGAPARSPSPSTQPWEHVNAIQE ARRLLNLSRDTAAEMNETVEVISEMFDLQEPTCLQTRLELYKQGLRGSLTKLKGPLTMM ASHYKQHCPPTPETSCATQIITFESFKENLKDFLLVIPFDCWEPVQEGSAGSAAGSGARR GVRVAWREPGRMELNMPHGQE (SEQ ID NO: 40);METDTLLLWVLLLWVPGSTGREEFEEEFEHREEFEHENLYFQGARRGVHVGWREPGRMELN MPHGGAEAAAKEAAAKAGAPARSPSPSTQPWEHVNAIQEARRLLNLSRDTAAEMNET VEVISEMFDLQEPTCLQTRLELYKQGLRGSLTKLKGPLTMMASHYKQHCPPTPETSCAT QIITFESFKENLKDFLLVIPFDCWEPVQE (SEQ ID NO: 36); andRRGVHVGWREPGRMELNMPHGGAEAAAKEAAAKAGAPARSPSPSTQPWEHVNAIQEA ...

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Номер записи: 22
20-01-2022 дата публикации

IMPROVED VECTOR SYSTEMS FOR CAS PROTEIN AND SGRNA DELIVERY, AND USES THEREFOR

Номер: US20220017921A1
Автор: Doench John, Dubrot Juan, Haining William Nicholas, Manguso Robert, Yates Kathleen
Принадлежит:

The present disclosure provides vectors, methods and kits for for delivery and stable expression of CRISPR/Cas components capable of inducing genetic modification of cells, followed by recombinase-mediated excision of some or all of these components after the cells have been successfully genetically modified. The disclosed vectors and methods provide for reduced immunogenic effects arising from one or more CRISPR/Cas components. The disclosed vectors comprise coding sequences that encode a Cas protein, detectable markers and a guide RNA. The disclosed vectors provide for the subsequent genomic excision of the CRISPR/Cas components after successful genetic modification, as mediated by recombinase recognition of recombination sites flanking one or more of the disclosed coding sequences. The present disclosure further provides methods of generating a population of genetically modified tumor cells for screening a candidate target gene for cancer immunotherapy. 1. A method of producing a population of genetically modified cells , comprising:(i) providing a population of cells; wherein the first integration vector is a replication defective retroviral vector derived from a primate lentivirus,', 'wherein the first integration vector comprises a first nucleic acid sequence comprising a first promoter operably linked to a Cas protein coding sequence encoding a Cas protein; and at least a first 3′ site-specific recombination site located 3′ to the Cas coding sequence, and', 'wherein the first integrating vector is capable of integration into the genomes of at least a portion of the population of cells;, '(ii) introducing a first integration vector into at least a portion of the population of cells,'}(iii) introducing an sgRNA into at least a portion of the population of cells, wherein the sgRNA is capable of guiding the Cas protein to a target site in the genomes of at least a portion of the population of cells, and wherein the Cas protein is capable of double-stranded DNA ...

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Номер записи: 23
14-01-2016 дата публикации

Animal Models of Cancer

Номер: US20160007578A1
Автор: Rogers Christopher, Swart John
Принадлежит: Exemplar Genetics, LLC

The present invention provides transgenic, large non-human animal models of cancer, as well as methods of using such animal models in the identification and characterization of therapies for cancer. 1185-. (canceled)186. A transgenic animal that models a human disease or condition , comprising a large , non-human transgenic animal comprising a p53 mutation wherein the p53 mutation comprises a mutation of an endogenous p53 gene and wherein the p53 mutation results in an altered expression of a p53 translation product and/or in expression of a non-functional p53 protein that corresponds to an alteration in a human p53 gene associated with a human disease or condition.187. The transgenic animal of claim 186 , wherein said transgenic animal is an ungulate.188. The transgenic animal of claim 187 , wherein said ungulate is selected from the group consisting of a swine claim 187 , a cows claim 187 , a sheep claim 187 , and a goats.189. The transgenic animal of claim 188 , wherein said ungulate is a swine that models a human cancer.190. The transgenic animal of claim 186 , wherein said mutation is in exon 5 of the endogenous p53 gene.191. The transgenic animal of claim 186 , wherein said p53 mutation is a gain of function mutation.192. The transgenic animal of claim 190 , wherein said p53 mutation results in an R167H substitution.193. The transgenic animal of claim 186 , wherein said p53 mutation is present in both alleles of the p53 gene.194. The transgenic animal of claim 186 , wherein said p53 mutation is a missense mutation resulting in a substitution of an amino acid in the endogenous p53 selected from Y220 claim 186 , G245 claim 186 , R248 claim 186 , and R273.195. The transgenic animal of claim 186 , wherein said p53 mutation is a deletion claim 186 , a disruption claim 186 , or an activation of both alleles of the endogenous p53 gene.196. The transgenic animal of claim 186 , wherein the transgenic animal further comprises a KRAS mutation claim 186 , wherein said ...

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Номер записи: 24
11-01-2018 дата публикации

Optimised Coding Sequence and Promoter

Номер: US20180008726A1
Автор: Davidoff Andrew, Gray John, McVey John, Nathwani Amit, Thrasher Adrian, Tuddenham Edward, Ward Natalie
Принадлежит:

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 ...

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Номер записи: 25
10-01-2019 дата публикации

AADC POLYNUCLEOTIDES FOR THE TREATMENT OF PARKINSON'S DISEASE

Номер: US20190008931A1
Автор: Kells Adrian Philip, Kotin Robert, Ravina Bernard
Принадлежит:

The disclosure relates to compositions and methods for the preparation, manufacture and therapeutic use of polynucleotides encoding AADC for the treatment of Parkinson's Disease. 1. An AAV vector genome comprising , in order:a) a 5′ inverted terminal repeat (ITR), wherein said 5′ ITR is 141 nucleotides in length;b) an AADC sequence region, said AADC sequence region comprising a nucleotide sequence encoding SEQ ID NO: 1,c) a 3′ ITR, wherein said 3′ ITR is 141 nucleotides in length.2. The AAV vector genome of claim 1 , wherein the 5′ ITR and the 3′ ITR are derived from AAV2.3. The AAV vector genome of claim 2 , wherein the AAV vector genome comprises a cytomegalovirus (CMV) sequence region derived from a CMV gene; wherein the CMV sequence region comprises an enhancer region and promoter region.4. The AAV vector genome of claim 3 , wherein the CMV sequence region is 507 nucleotides in length.5. The AAV vector genome of claim 2 , wherein the AAV vector genome comprises an immediate early 1 (IE1) sequence region derived from an IE1 gene.6. The AAV vector genome of claim 5 , wherein the IE1 sequence region comprises a nucleotide sequence from IE1 exon1.7. The AAV vector genome of claim 6 , wherein the IE1 sequence region comprises a nucleotide sequence from IE1 intron 1 or a fragment thereof.8. The AAV vector genome of claim 7 , wherein the IE1 sequence region is 166 nucleotides in length.9. The AAV vector genome of claim 2 , wherein the AAV vector genome comprises a human beta globin (BB) sequence region derived from a HB gene.10. The AAV vector genome of claim 9 , wherein the HB sequence region comprises a nucleotide sequence from HB intron 2.11. The AAV vector genome of claim 10 , wherein the HB sequence region comprises a nucleotide sequence from HB exon 3.12. The AAV vector genome of claim 11 , wherein the HB sequence region is 400 nucleotides in length.13. The AAV vector genome of claim 2 , wherein the AAV vector genome comprises a poly(A) signal sequence region ...

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Номер записи: 26
10-01-2019 дата публикации

Aadc polynucleotides for the treatment of parkinson's disease

Номер: US20190008932A1
Автор: Adrian Philip KELLS, Bernard RAVINA, Robert Kotin
Принадлежит: Voyager Therapeutics Inc

The disclosure relates to compositions and methods for the preparation, manufacture and therapeutic use of polynucleotides encoding AADC for the treatment of Parkinson's Disease.

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Номер записи: 27
10-01-2019 дата публикации

AADC POLYNUCLEOTIDES FOR THE TREATMENT OF PARKINSON'S DISEASE

Номер: US20190008933A1
Автор: Kells Adrian Philip, Kotin Robert, Ravina Bernard
Принадлежит:

The disclosure relates to compositions and methods for the preparation, manufacture and therapeutic use of polynucleotides encoding AADC for the treatment of Parkinson's Disease. 1. An AAV vector genome comprising , in order:a) a 5′ inverted terminal repeat (ITR), wherein said 5′ ITR is 119 nucleotides in length;b) an AADC sequence region, said AADC sequence region comprising a nucleotide sequence encoding SEQ ID NO: 1,c) a 3′ ITR, wherein said 3′ ITR is 130 nucleotides in length.2. The AAV vector genome of claim 1 , wherein the 5′ ITR and the 3′ ITR are derived from AAV2.3. The AAV vector genome of claim 2 , wherein the 5′ ITR consists of a nucleotide sequence consisting of nucleotides 3535 to 3417 of SEQ ID NO 6.4. The AAV vector genome of claim 2 , wherein the 5′ ITR consists of a nucleotide sequence which has at least 99% sequence identity with nucleotides 3535 to 3417 of SEQ ID NO 6.5. The AAV vector genome of claim 3 , wherein the 3′ ITR consists of a nucleotide sequence consisting of nucleotides 130 to 1 of SEQ ID NO 6.6. The AAV vector genome of claim 4 , wherein the 3′ ITR consists of a nucleotide sequence which has at least 99% sequence identity with nucleotides 130 to 1 of SEQ ID NO 6.7. The AAV vector genome of claim 6 , wherein the AAV vector genome comprises a cytomegalovirus (CMV) sequence region derived from a CMV gene; wherein the CMV sequence region comprises an enhancer region and promoter region.8. The AAV vector genome of claim 7 , wherein the CMV sequence region comprises a nucleotide sequence which has at least 99% sequence identity with nucleotides 263 to 769 of SEQ ID NO 6.9. The AAV vector genome of claim 7 , wherein the CMV sequence region comprises a nucleotide sequence which has at least 98% sequence identity with nucleotides 263 to 769 of SEQ ID NO 6.10. The AAV vector genome of claim 8 , wherein the CMV enhancer region comprises a nucleotide sequence which has at least 99% sequence identity with nucleotides 263 to 566 of SEQ ID NO 6.11 ...

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Номер записи: 28
10-01-2019 дата публикации

COMPUTATIONALLY OPTIMIZED BROADLY REACTIVE ANTIGENS FOR INFLUENZA

Номер: US20190008949A1
Автор: Giles Brendan M., Ross Ted M.
Принадлежит: University of Pittsburgh - of the Commonwealth System of Higher Education

The development of a computationally optimized influenza HA protein that elicits broadly reactive immune response to all H5N1 influenza virus isolates is described. The optimized HA protein was developed through a series of HA protein alignments, and subsequent generation of consensus sequences, for clade 2 H5N1 influenza virus isolates. The final consensus HA amino acid sequence was reverse translated and optimized for expression in mammalian cells. Influenza virus-like particles containing the optimized HA protein are an effective vaccine against H5N1 influenza virus infection in animals. 1. A method of eliciting a broadly reactive immune response against influenza virus in a subject , comprising (i) obtaining the amino acid sequences of the polypeptide from a group of influenza virus isolates, wherein the influenza virus isolates are from the same subtype;', '(ii) organizing the amino acid sequences of the polypeptide from the group of influenza virus isolates by clade or sub-clade and then by geographical region within each clade or sub-clade;', '(iii) aligning the amino acid sequences within each geographical region to generate primary consensus sequences, wherein each geographic region is represented by a primary consensus sequence;', '(iv) aligning the primary consensus sequences to generate secondary consensus sequences, wherein each clade or sub-clade is represented by a secondary consensus sequence; and', '(v) aligning the secondary consensus sequences, thereby generating the optimized influenza virus polypeptide sequence; and, 'generating an optimized influenza virus polypeptide sequence comprising the steps ofadministering the optimized influenza virus polypeptide to the subject.2. The method of claim 1 , wherein generating the optimized influenza virus polypeptide sequence further comprises:(vi) reverse translating the optimized influenza virus polypeptide sequence to generate a coding sequence; and(vii) optimizing the coding sequence for expression in ...

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Номер записи: 29
11-01-2018 дата публикации

COMPOSITIONS AND METHODS FOR MONITORING IN REAL-TIME CONSTRUCTION AND BIOENGINEERING OF MAMMALIAN SYNTHETIC CHROMOSOMES

Номер: US20180010150A1
Автор: Greene Amy, Perkins Edward
Принадлежит:

The present invention encompasses compositions and methods to allow one to monitor formation of synthetic chromosomes in real-time via standardized fluorescent technology, eliminating the need for cumbersome, expensive, and possibly mutagenic analysis. 1. A method for screening production of synthetic chromosomes comprising:providing a synthetic chromosome production reporter cell line;transfecting the synthetic chromosome production reporter cell line with an endogenous chromosome tag and a synthetic chromosome tag;transfecting the synthetic chromosome production reporter cell line with synthetic chromosome production components; andmonitoring production of the synthetic chromosome in the synthetic chromosome production reporter cell line by using the endogenous chromosome tag and the synthetic chromosome tag.2. The method of claim 1 , wherein the endogenous chromosome tag and a synthetic chromosome tag are stably integrated into the genome of the synthetic chromosome production reporter cell line.3. The method of claim 1 , wherein the endogenous chromosome tag becomes stably integrated into the genome of the synthetic chromosome production reporter cell line and the synthetic chromosome tag becomes stably integrated into the synthetic chromosome.4. The method of claim 1 , wherein the endogenous chromosome tag and the synthetic chromosome tag are stably integrated into the synthetic chromosome.5. The method of claim 1 , wherein the endogenous chromosome tag becomes stably integrated into the synthetic chromosome and the synthetic chromosome tag becomes stably integrated into the genome of synthetic chromosome production reporter cell line.6. The method claim 1 , wherein an arm of an endogenous chromosome present in the synthetic chromosome production reporter cell line comprises a recombination site compatible for interaction with a recombination site in the synthetic chromosome.7. The method claim 1 , wherein the endogenous chromosome tag and the synthetic ...

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Номер записи: 30
14-01-2021 дата публикации

FOOD COMPOSITIONS COMPRISING MILK PROTEINS PRODUCED IN TRANSGENIC PLANTS

Номер: US20210010017A1
Автор: EL-RICHANI Magi, Li Shu
Принадлежит:

The disclosure describes a transgenic dicot or monocot plant having bovine milk protein(s) and methods of producing the transgenic dicot or monocot plant containing bovine milk protein(s). These transgenic dicot or monocot plants can express and produce bovine milk protein(s). The methods involve introducing a recombinant DNA construct expressing a bovine milk protein into a dicot or monocot plant, obtaining the dicot or monocot plant containing the bovine milk protein(s) from a recombinant DNA construct, cultivating and harvesting the transgenic dicot or monocot plant, and extracting and purifying the bovine milk protein(s) from transgenic dicot or monocotyledonous plants. The disclosure also describes food compositions comprising milk proteins produced using the transgenic dicot or monocot plants described herein. 1. A food composition comprising a milk protein , wherein the milk protein is produced in a transgenic plant , and wherein the milk protein is α-S1 casein , α-S2 casein , β-casein , κ-casein , α-lactalbumin , β-lactoglobulin , or lysozyme.2. The food composition of claim 1 , wherein the milk protein is κ-casein.3. The food composition of claim 2 , wherein the κ-casein has a sequence with at least 90% sequence identity to SEQ ID NO: 5.4. The food composition of claim 2 , wherein the κ-casein has the sequence of SEQ ID NO: 5.5. The food composition of claim 1 , wherein the milk protein has a sequence with at least 90% sequence identity to any one of SEQ ID NO: 7 claim 1 , SEQ ID NO: 11 claim 1 , SEQ ID NO:12 claim 1 , SEQ ID NO:22 claim 1 , SEQ ID NO:23 claim 1 , or SEQ ID NO:24.6. The food composition of claim 1 , wherein the transgenic plant is a dicot.7Arabidopsis. The food composition of claim 6 , wherein the transgenic plant is claim 6 , tobacco claim 6 , tomato claim 6 , potato claim 6 , sweet potato claim 6 , cassava claim 6 , carrot claim 6 , strawberry claim 6 , lettuce claim 6 , oak claim 6 , maple claim 6 , walnut claim 6 , rose claim 6 , mint ...

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Номер записи: 31
14-01-2021 дата публикации

Viral and non-viral nanoplasmid vectors with improved production

Номер: US20210010021A1
Автор: James A. Williams
Принадлежит: Nature Technology Corp

A method for improving the replication of a covalently closed circular plasmid is provided. The method includes providing a covalently closed circular plasmid having a Pol I-dependent origin of replication, and an insert including a structured DNA sequence selected from the group consisting of inverted repeat sequence, direct repeat sequence, homopolymeric repeat sequence, eukaryotic origin of replication or eukaryotic promoter enhancer sequence, wherein the structured DNA sequence is located at a distance of less than 1000 by from the Pol I-dependent origin of replication in the direction of replication. The method also includes modifying the covalently closed circular recombinant molecule such that the Poi I-dependent origin of replication is replaced with a Pol III-dependent origin of replication, whereby the resultant Pol III-dependent origin of replication covalently closed circular plasmid has improved replication. An antibiotic marker free covalently closed circular recombinant DNA molecule is also provided.

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Номер записи: 32
14-01-2021 дата публикации

TREATMENT OF OCULAR DISEASES WITH HUMAN POST-TRANSLATIONALLY MODIFIED VEGF-TRAP

Номер: US20210010025A1
Автор: Danos Olivier, Gerner Franz Michael, Van Everen Sherri, Wu Zhuchun
Принадлежит:

Compositions and methods are described for the delivery of a fully human post-translationally modified (HuPTM) therapeutic VEGF-Trap (VEGF-Trap)—to a human subject diagnosed with an ocular disease or condition or cancer associated with neovascularization and indicated for treatment with the therapeutic mAb. Delivery may be advantageously accomplished via gene therapy—e.g., by administering a viral vector or other DNA expression construct encoding the VEGF-Trapto a patient (human subject) diagnosed with an ocular condition or cancer indicated for treatment with the VEGF-Trap—to create a permanent depot in a tissue or organ of the patient that continuously supplies the VEGF-Trap, i.e., a human-glycosylated transgene product. Alternatively, the VEGF-Trap, for example, produced in cultured human cell culture, can be administered to the patient for treatment of the ocular disease or cancer. 1. An expression construct comprising an expression cassette flanked by AAV inverted terminal repeats (ITRs) wherein the expression cassette comprises a transgene encoding a VEGF-TrapHuPTM operably linked to one or more regulatory sequences that control expression of the transgene in human retinal cells or human liver cells , wherein the transgene encodes a leader sequence operable in human retinal cells or human liver cells and a VEGF-TrapHuPTM , wherein the VEGF-TrapHuPTM comprises an amino acid sequence having amino acid residues 1 to 204 of SEQ ID NO: 1.2. The expression construct of wherein the VEGF-TrapHuPTM comprises an amino acid sequence having amino acid residues 1 to 205 of SEQ ID NO: 1 linked at the C terminus to an IgG1 claim 1 , IgG2 claim 1 , or IgG4 Fc region comprising at least a partial hinge region at the N-terminus of the Fc region.3. The expression construct of claim 2 , wherein the Fc region comprises a full hinge region.4. The expression construct of claim 2 , wherein one or more of the cysteine residues within the hinge region is substituted with a serine.5. ...

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Номер записи: 33
09-01-2020 дата публикации

COMPOSITIONS AND METHODS FOR ENHANCED GENE EXPRESSION

Номер: US20200010851A1
Автор: KERAVALA Annahita
Принадлежит:

The present disclosure provides polynucleotide cassettes, expression vectors and methods for the expression of a gene in mammalian cells. 1. A non-naturally occurring polynucleotide cassette for enhanced expression of a transgene in a mammalian cell , comprising in 5′ to 3′ order:(a) a first enhancer region;(b) a promoter region;(c) a coding sequence encoding a secretory polypeptide;(d) a second enhancer region; and(e) a polyadenylation site,wherein the coding sequence is operably linked to the promoter region,wherein the expression of the secretory polypeptide from the polynucleotide cassette in mammalian cells is at least 5× higher than the expression of the secretory polypeptide from a reference cassette in the mammalian cells, andwherein the reference cassette comprises, in 5′ to 3′ order, a CMV enhancer sequence (SEQ ID NO:2), a CMV promoter (SEQ ID NO:21), a chimeric intron (SEQ ID NO:22), a 5′UTR (SEQ ID NO:23), a coding sequence encoding the secretory polypeptide, a 3′UTR (SEQ ID NO:25), and an SV40 polyA sequence (SEQ ID NO:26).2. The polynucleotide cassette of claim 1 , wherein the cassette does not contain an RNA export signal.3. The polynucleotide cassette of any of the preceding claims claim 1 , wherein the first enhancer region comprises the cytomegalovirus (CMV) sequence set forth in SEQ ID NO:1 or a sequence with at least 85% identity thereto.4. The polynucleotide cassette of any of the preceding claims claim 1 , wherein the promoter region comprises the CMV promoter sequence set forth in SEQ ID NO:4 or a sequence with at least 85% identity thereto.5. The polynucleotide cassette of any of the preceding claims claim 1 , further comprising an untranslated region (5′UTR) downstream of the promoter region and upstream of the coding sequence claim 1 , wherein the 5′UTR comprises claim 1 , in 5′ to 3′ order claim 1 , a TPL sequence according to SEQ ID NO:11 or a sequence with at least 85% identity thereto claim 1 , and an eMLP sequence according to SEQ ID ...

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Номер записи: 34
03-02-2022 дата публикации

ADENO-ASSOCIATED VIRAL VECTORS FOR THE TREATMENT OF BEST DISEASE

Номер: US20220033826A1
Автор: Hauswirth William W., Ildefonso Cristhian J., Lewin Alfred S., Young Brianna M.
Принадлежит: University of Florida Research Foundation, Incorporated

Aspects of the disclosure relate to methods and compositions useful for treating bestrophinopathies, such as Best Disease. 1. A short hairpin RNA (shRNA) comprising:a) a sense strand comprising the nucleotide sequence CGUCAAAGCUUCACAGUGU (SEQ ID NO: 2) and an antisense strand comprising the nucleotide sequence ACACUGUGAAGCUUUGACG (SEQ ID NO: 3); andb) a loop.2. The shRNA of claim 1 , wherein the loop comprises the nucleotide sequence UUCAAGAGA (SEQ ID NO: 7).3. The shRNA of claim 1 , wherein the shRNA comprises the nucleotide sequence CGUCAAAGCUUCACAGUGUUUCAAGAGAACACUGUGAAGCUUUGACG (SEQ ID NO: 1).4. A vector encoding the shRNA of .5. The vector of further comprising a recombinant bestrophin (BEST1) coding sequence that does not contain a sequence targeted by the shRNA.6. The vector of claim 5 , wherein the recombinant BEST1 coding sequence is codon-optimized for expression in a human cell.7. The vector of claim 5 , wherein the recombinant BEST1 coding sequence comprises a nucleotide sequence that is at least 90% identical to the nucleotide sequence of SEQ ID NO: 9.8. The vector of claim 7 , wherein the recombinant BEST1 coding sequence comprises the nucleotide sequence of SEQ ID NO: 9.9. A vector encoding an shRNA of and a recombinant BEST1 sequence comprising a nucleotide sequence that is at least 90% identical to the nucleotide sequence of SEQ ID NO: 11.10. The vector of claim 9 , wherein the vector comprises the nucleotide sequence of SEQ ID NO: 11.11. The vector of claim 4 , wherein the vector is a plasmid or a viral vector.12. (canceled)13. The vector of claim 11 , wherein the viral vector is a recombinant adeno-associated viral (rAAV) vector.14. The vector of claim 13 , wherein the rAAV vector is self-complementary.15. A recombinant adeno-associated viral (rAAV) particle comprising the rAAV vector of .16. The rAAV particle of claim 15 , wherein the rAAV viral particle is an AAV serotype 2 (AAV2) viral particle.17. A composition comprising the rAAV particle of ...

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Номер записи: 35
15-01-2015 дата публикации

COMBINATION MEDICAMENT COMPRISING IL-12 AND AN AGENT FOR BLOCKADE OF T-CELL INHIBITORY MOLECULES FOR TUMOUR THERAPY

Номер: US20150017121A1
Автор: Becher Burkhard, VOM BERG Johannes
Принадлежит: UNIVERSITAT ZURICH

The invention relates to a combination medicament for treatment of malignant neoplastic disease. The combination medicament comprises an IL-12 polypeptide having a biological activity of IL-12 or a nucleic acid expression vector comprising a sequence encoding such IL-12 polypeptide, and a non-agonist CTLA-4 ligand or non-agonist PD-1 ligand, particularly an anti-CTLA-4 or anti-PD-1 immunoglobulin G. 114.-. (canceled)15. A method of treating a patient suffering from malignant neoplastic disease , comprising the administration into a tumour , into the vicinity of a tumour , or to the lymph node associated with a tumour , of an IL-12 polypeptide having a biological activity of IL-12 or a nucleic acid expression vector encoding said IL-12 polypeptide , and the administration of a T cell inhibition blocker agent selected from a non-agonist CTLA-4 ligand and a non-agonist PD-1 ligand.16. A polypeptide comprisinga. a polypeptide sequence at least 95% identical to the sequence of human p35 (SEQ ID 05), andb. a polypeptide sequence at least 95% identical to the sequence of human p40 (SEQ ID 06) andc. a human immunoglobulin G subgroup 4 crystallisable fragment.17. The polypeptide of claim 16 , having a sequence at least 95% identical to SEQ ID 01.1820.-. (canceled)21. The method of claim 15 , wherein the IL-12 polypeptide comprises:a. a polypeptide sequence at least 95% identical to the sequence of human p35 (SEQ ID 05), andb. a polypeptide sequence at least 95% identical to the sequence of human p40 (SEQ ID 06).22. The method of claim 21 , wherein the IL-12 polypeptide comprises an immunoglobulin G crystallisable fragment.23. The method of claim 21 , wherein the IL-12 polypeptide comprises a human immunoglobulin G subgroup 4 crystallisable fragment.24. The method of claim 23 , wherein the IL-12 polypeptide comprisesa. an immunoglobulin G crystallisable fragment and a recombinant or synthetic human IL-12 sequence, orb. a sequence at least 95% identical to SEQ ID 01.25. The ...

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Номер записи: 36
17-01-2019 дата публикации

Multimerization of Recombinant Protein by Fusion to a Sequence from Lamprey

Номер: US20190016760A1
Автор: Legastelois Isabelle, Sodoyer Régis
Принадлежит: Sanofi Pasteur

The present invention relates to polymerized recombinant proteins, to recombinant nucleic acids coding for the polymerized recombinant proteins, to expression cassettes comprising the recombinant nucleic acids, to host cells transformed by the expression cassettes and to a method for multimerizing a recombinant protein. The polymerized proteins of the invention may be used in pharmaceutical or immunogenic compositions. In particular, the recombinant proteins may be antigens, antibodies or scaffolds. In particular, the polymerized recombinant protein may be an influenza haemagglutinin. 1. A molecule which comprises a first amino acid sequence which has at least 80% identity to SEQ ID NO: 1 and a second amino acid sequence which is heterologous to said first sequence , wherein said molecule does not comprise a leucine-rich repeat (LRR) module from a lamprey VLR-B antibody.2. A molecule according to claim 1 , wherein said molecule does not comprise a sequence selected from the group of sequences defined by SEQ ID NO: 29.3. A molecule according to or claim 1 , wherein the only amino acid sequence in said molecule which is derived from a lamprey VLR-B antibody is the sequence having at least 80% identity to SEQ ID NO: 1.4. The molecule according to any one of to wherein said molecule is a recombinant protein.5. The molecule according to any one of to which comprises cysteine residues at the positions within the molecule corresponding to positions 2 claim 1 , 7 claim 1 , 13 claim 1 , 19 claim 1 , 21 claim 1 , 24 and 27 of SEQ ID NO:1.6. The molecule according to any one of to wherein the first amino acid sequence has at least 90% identity or 100% identity to SEQ ID NO: 1.7. The molecule according to any one of to which comprises SEQ ID NO: 2.8. The molecule according to any one of to claim 1 , wherein there is a linker between the first amino acid sequence and the heterologous amino acid sequence.9. The molecule according to any one of to claim 1 , wherein the ...

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Номер записи: 37
16-01-2020 дата публикации

MEANS FOR GENERATING ADENOVIRAL VECTORS FOR CLONING LARGE NUCLEIC ACIDS

Номер: US20200017863A1
Автор: Thirion Christian
Принадлежит:

The present invention is related to a nucleic acid molecule, which is also referred to as third nucleic acid molecule, wherein the third nucleic acid molecule comprises 1. A nucleic acid molecule , which is also referred to as third nucleic acid molecule , wherein the third nucleic acid molecule comprises (a) optionally, a first part of a genome of a virus;', '(b) a nucleotide sequence, preferably a genomic nucleotide sequence, or a transcription unit;', '(c) a regulatory nucleic acid sequence which has a regulatory activity in a prokaryote;', '(d) a site-specific recombination site;', '(e) a nucleotide sequence providing for a negative selection marker;', '(f) a bacterial nucleotide sequence unit comprising (i) bacterial nucleotide sequences for conditional replication and (ii) a nucleotide sequence providing for a positive selection marker; and', '(g) optionally a first restriction site; or, '(1) a nucleic acid molecule comprising the following elements(2) a nucleic acid molecule comprising a nucleotide sequence according to SEQ ID NO: 6; or(3) a nucleic acid molecule identical or similar to the nucleic acid molecule contained in the organism deposited with the DSMZ under the Budapest treaty under accession number DSM 23754, wherein preferably the nucleic acid molecule contained in the organism is a heterologous nucleic acid molecule;wherein the third nucleic acid molecule is either a linear or a circular molecule.2. The third nucleic acid molecule according to claim 1 , wherein in the nucleic acid molecule of (1) the regulatory nucleic acid sequence which has a regulatory activity in a prokaryote claim 1 , the site-specific recombination site and the nucleotide sequence providing for a negative selection marker are arranged in a 5′ to 3′ direction.3. The third nucleic acid molecule according to any one of to claim 1 , wherein the third nucleic acid molecule contains exactly one site-specific recombination site.4. The third nucleic acid molecule according to any ...

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Номер записи: 38
21-01-2021 дата публикации

Genome Editing without Nucleases

Номер: US20210017539A1
Автор: Barzel Adi, Kay Mark A.
Принадлежит:

Methods and compositions are provided for editing the genome of a cell without the use of an exogenously supplied nuclease. Aspects of the methods include contacting a cell with a targeting vector comprising nucleic acid sequence to be integrated into the target locus, where the cell is not also contacted with a nuclease. In addition, reagents, devices and kits thereof that find use in practicing the subject methods are provided. 1. A method for the targeted integration of a transgene into the genome of a cell in the absence of an exogenously provided nuclease , the method comprising:contacting a cell with a recombinant viral vector, the recombinant viral vector comprising:i. a polynucleotide comprising a first nucleic acid sequence and a second nucleic acid sequence, wherein the first nucleic acid sequence encodes the transgene; and the second nucleic acid sequence is positioned 5′ or 3′ to the first nucleic acid sequence and promotes the production of two independent gene products upon integration into the target integration site in the genome of the cell;ii. a third nucleic acid sequence positioned 5′ to the polynucleotide and comprising sequence that is substantially homologous to genomic sequence 5′ of a target integration site in the genome of the cell; andiii. a fourth nucleic acid sequence positioned 3′ of the polynucleotide and comprising sequence that is substantially homologous to genomic sequence 3′ of a target integration site in the genome of the cell;wherein the cell is not contacted with a nuclease or nucleic acid encoding a nuclease.2. The method according to claim 1 , wherein the cell is a non-dividing cell.3. The method according to claim 1 , wherein the contacting occurs in vivo.4. The method according to claim 3 , wherein the method finds use in treating a medical condition associated with a gene deficiency.5. The method according to claim 4 , wherein the medical condition is selected from the group consisting of hemophilia claim 4 , hemophilia ...

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Номер записи: 39
24-01-2019 дата публикации

COMMON LIGHT CHAIN MOUSE

Номер: US20190021295A1
Автор: BABB Robert, BUCKLER David R., DAVIS Samuel, LEVENKOVA Natasha, MACDONALD Lynn, MCWHIRTER John, MEAGHER Karolina A., MURPHY Andrew J., STEVENS Sean
Принадлежит:

A genetically modified mouse is provided, wherein the mouse expresses an immunoglobulin light chain repertoire characterized by a limited number of light chain variable domains. Mice are provided that express just one or a few immunoglobulin light chain variable domains from a limited repertoire in their germline. Methods for making bispecific antibodies having universal light chains using mice as described herein, including human light chain variable regions, are provided. Methods for making human variable regions suitable for use in multispecific binding proteins, e.g., bispecific antibodies, and host cells are provided. Bispecific antibodies capable of binding first and second antigens are provided, wherein the first and second antigens are separate epitopes of a single protein or separate epitopes on two different proteins are provided. 1. A mouse that expresses a single rearranged human immunoglobulin light chain variable (V) region derived from a rearranged germline variable region sequence that comprises a human Vgene segment derived from a human Vκ3-20 gene segment and{'sub': H', 'L', 'H', 'H', 'H', 'H', 'H', 'H', 'H', 'H', 'H', 'H', 'H', 'H', 'H', 'H', 'H', 'H', 'H', 'H', 'H', 'H', 'H', 'H', 'H', 'H', 'H', 'H', 'H', 'H', 'H', 'H', 'H', 'H', 'H', 'H', 'H', 'H', 'H', 'H', 'H, 'a human immunoglobulin heavy chain variable (V) region cognate with said Vregion wherein the Vregion comprises a human Vsegment derived from a segment selected from V6-1, V1-2, V1-3, V2-5, V3-7, V1-8, V3-9, V3-11, V3-13, V3-15, V1-18, V3-20, V3-21, V3-23, V1-24, V2-26, V4-28, V3-30, V4-31, V3-33, V4-34, V4-39, V3-43, V1-46, V3-48, V3-49, V5-51, V3-53, V1-58, V4-59, V4-61, V3-64, V3-66, V1-69, V2-70, V3-72, and V3-73.'}2. The mouse of claim 1 , wherein the Vregion comprises a human Vsegment derived from a segment selected from V1-2 claim 1 , V1-3 claim 1 , V2-5 claim 1 , V3-7 claim 1 , V1-8 claim 1 , V3-9 claim 1 , V3-13 claim 1 , V3-15 claim 1 , V3-20 claim 1 , V3-21 claim 1 , V3-23 ...

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Номер записи: 40
25-01-2018 дата публикации

METHOD FOR INDUCING TARGETED MEIOTIC RECOMBINATIONS

Номер: US20180023091A1
Автор: BASTIANELLI GIACOMO, NICOLAS ALAIN, SERERO ALEXANDRE
Принадлежит:

The present invention relates to a fusion protein comprising a Cas9 domain and a Spo11 domain, as well as the use of this protein to induce targeted meiotic recombinations in a eukaryotic cell. 127-. (canceled)28. A method for inducing targeted meiotic recombinations in a eukaryotic cell comprising:introducing into said cell:a) a fusion protein comprising a Cas9 domain and a Spo11 domain, or a nucleic acid encoding said fusion protein; andb) one or more guide RNAs or one or more nucleic acids encoding said guide RNAs, said guide RNAs comprising an RNA structure for binding to the Cas9 domain of the fusion protein and a sequence complementary to the targeted chromosomal region; andinducing said cell to enter meiotic prophase I.29. The method according to claim 28 , wherein the fusion protein further comprises a nuclear localization signal sequence.30. The method according to claim 28 , wherein the Cas9 domain is a nuclease-deficient Cas9 protein.31. The method according to claim 28 , wherein the expression of the nucleic acid encoding said fusion protein is placed under the control of a constitutive claim 28 , inducible or meiosis-specific promoter.32. The method according to claim 28 , further comprising introducing one or more additional guide RNAs targeting one or more other chromosomal regions claim 28 , or nucleic acids encoding said additional guide RNAs.33. The method according to claim 28 , wherein the cell is a yeast.34. The method according to claim 28 , wherein the eukaryotic cell is a plant cell.35. A fusion protein comprising a Cas9 domain and a Spo11 domain.36. A nucleic acid encoding the fusion protein according to .37. An expression cassette or a vector comprising the nucleic acid according to .38. The vector according to claim 37 , said vector being a plasmid comprising:a bacterial origin of replication,an expression cassette comprising a nucleic acid encoding a fusion protein comprising a Cas9 domain and a Spo11 domain under the control of an ...

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Номер записи: 41
24-01-2019 дата публикации

VECTOR

Номер: US20190024116A1
Автор: Kokaia Merab
Принадлежит:

The present invention relates to a recombinant adeno-associated viral (r AAV) vector comprising neuropeptide Y (NPY) coding sequence and neuropeptide Y2 receptor (NPY2R) coding sequence. The invention further relates to a AAV particle comprising said vector, wherein the vector is encapsulated by adeno-associated virus (AAV) capsid proteins. Also, a pharmaceutical composition comprising said AAV particle, for use in the prevention or treatment of a neurological disorder in mammals, such as epilepsy. 1. A recombinant adeno-associated viral (rAAV) vector comprising a neuropeptide Y (NPY) coding sequence and a neuropeptide Y2 receptor (NPY2R) coding sequence.247-. (canceled)48. The vector according to claim 1 , wherein said neuropeptide Y (NPY) coding sequence comprises a sequence corresponding to SEQ ID NO:1 or a sequence having at least 90% sequence identity to SEQ ID NO:1.49. The vector according to claim 1 , wherein said neuropeptide Y2 receptor (NPY2R) coding sequence comprises a sequence corresponding to SEQ ID NO:2 or a sequence having at least 90% sequence to SEQ ID NO:2.50. The vector according to claim 1 , wherein the vector further comprises at least one of the functional elements of AAV2 Inverted Terminal Repeat sequences (ITR) claim 1 , hybrid cytomegalovirus enhancer/chicken beta-actin CAG promoter (CAG) claim 1 , internal ribosome entry site (IRES) claim 1 , woodchuck hepatitis post-translational regulatory element (WPRE) claim 1 , or bovine growth hormone polyadenylation (bGH-polyA) signal sequence.51. The vector according to claim 50 , wherein the vector comprises a hybrid cytomegalovirus enhancer/chicken beta-actin CAG promoter (CAG) and said CAG promoter sequence is located upstream of the coding sequences for NPY and NPY2R.52. The vector according to claim 50 , wherein the vector comprises an internal ribosome entry site (IRES) and said IRES sequence is located between the coding sequences for NPY and NPY2R.53. The vector according to claim 50 , ...

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Номер записи: 42
23-01-2020 дата публикации

MICE EXPRESSING A LIMITED IMMUNOGLOBULIN LIGHT CHAIN REPERTOIRE

Номер: US20200024368A1
Автор: MACDONALD Lynn, MCWHIRTER John, MURPHY Andrew J., STEVENS Sean
Принадлежит:

A genetically modified mouse is provided, wherein the mouse expresses an immunoglobulin light chain repertoire characterized by a limited number of light chain variable domains. Mice are provided that present a choice of two human light chain variable gene segments such that the immunoglobulin light chains expresses by the mouse comprise one of the two human light chain variable gene segments. Methods for making bispecific antibodies having universal light chains using mice as described herein, including human light chain variable regions, are provided. Methods for making human variable regions suitable for use in multispecific binding proteins, e.g., bispecific antibodies, and host cells are provided. 130-. (canceled)31. A method for generating a human immunoglobulin heavy or light chain variable region sequence comprising the steps of: 1. exactly two unrearranged human immunoglobulin Vκ gene segments and five unrearranged human immunoglobulin Jκ gene segments operably linked to a mouse immunoglobulin light chain constant region sequence at the endogenous kappa light chain loci of the mouse, wherein the two unrearranged human immunoglobulin Vκ gene segments are a human Vκ1-39 gene segment and a human Vκ3-20 gene segment; and', {'sub': H', 'H', 'H, '2. one or more unrearranged human immunoglobulin Vgene segments, one or more unrearranged human immunoglobulin Dgene segments, and one or more unrearranged human immunoglobulin Jgene segments operably linked to a mouse immunoglobulin heavy chain constant region sequence at the endogenous heavy chain loci of the mouse;'}], '(a) immunizing a genetically modified mouse with an antigen of interest, wherein the genetically modified mouse comprises in its germline genome{'sub': 'κ', 'wherein the unrearranged human immunoglobulin heavy chain and kappa light chain gene segments of the genetically modified mouse are capable of rearranging and encoding human immunoglobulin variable domains of an antibody, wherein the genetically ...

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Номер записи: 43
23-01-2020 дата публикации

Corynebacterium Constitutive Expression Vector Promoter Screened On The Basis Of Transcriptome Sequencing, Screening Method Thereof, And Applications Thereof

Номер: US20200024625A1
Автор: LI Jing, LIU Aifu, MEI Xuechen, SONG Mengjun, SU Haixia, WAN Kun, Wang Jiong, XING Panpan
Принадлежит:

Provided is a method for screening a constitutive expression vector promoter on the basis of transcriptome sequencing; and further provided are the constitutive expression vector promoter screened on the basis of transcriptome sequencing, an expression vector comprising the promoter, a recombination strain obtained by transforming a host cell using the expression vector, and applications thereof. 1Corynebacterium. A method for screening a constitutive expression vector promoter on the basis of transcriptome sequencing , comprising:{'i': 'Corynebacterium', 'analyzing the transcription level of each gene of the in logarithmic phase and stationary phase, and screening a class of genes with low transcription levels in the logarithmic phase and high transcription levels in the stationary phase are screened by analyzing the transcription abundance of each gene in two phases;'}amplifying the promoter fragments of genes and incorporating the DNA fragment of the promoter and marker gene into an expression vector to construct a promoter probe vector;transforming the probe vector into host cells, and carrying out continuous passage under an antibiotic-stress free condition;{'i': 'Corynebacterium', 'selecting host cells having low expression level of marker gene in the logarithmic phase and high expression level in the stationary phase, and continuously passaged for 50 generations without losing probe vector, thus the promotor transformed by the host cells is the constitutive expression vector promoter.'}2Corynebacterium. A constitutive expression vector promoter screened by the method of claim 1 , wherein the nucleotide sequence of the promoter is set forth in SEQ ID NO: 1 claim 1 , SEQ ID NO: 2 or SEQ ID NO: 3.3CorynebacteriumCorynebacterium. A constitutive expression vector comprising the constitutive expression vector promoter of .4CorynebacteriumCorynebacterium. The constitutive expression vector of claim 2 , wherein it is constructed by inserting a gene of interest and ...

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Номер записи: 44
04-02-2016 дата публикации

RECOMBINANT RSV WITH SILENT MUTATIONS, VACCINES, AND METHODS RELATED THERETO

Номер: US20160030549A1
Автор: Hotard Anne, Littauer Elizabeth, MENG Jia, Moore Martin L., Stobart Christopher
Принадлежит:

In certain embodiments, the disclosure relates to the polynucleotide sequences of respiratory syncytial virus (RSV). In certain embodiments, the disclosure relates to isolated or recombinant nucleic acids and polypeptides comprising desirable nucleic acid sequences and mutations disclosed herein. In certain embodiments, isolated or recombinant RSV comprising the nucleic acids and polypeptides disclosed herein (e.g., attenuated recombinant RSV) are also provided, as are immunogenic compositions including such nucleic acids, polypeptides, and RSV genomes that are suitable for use as vaccines. Attenuated or killed RSV containing these nucleic acids and mutation in the form of copied nucleic acids (e.g., cDNAs) are also contemplated. 1. An isolated recombinant nucleic acid encoding NS1 and/or NS2 of a wild-type human RSV or variant wherein the nucleotides are substituted such that a codon to produce Gly is GGT , a codon to produce Asp is GAT , a codon to produce Glu is GAA , a codon to produce His is CAT , a codon to produce Ile is ATA , a codon to produce Lys is AAA , a codon to produce Leu is CTA , a codon to produce Asn is AAT , a codon to produce Gln is CAA , a codon to produce Val is GTA , or a codon to produce Tyr is TAT , or combinations thereof.2. The isolated recombinant nucleic acid of further comprising a combination of at least two claim 1 , three claim 1 , four claim 1 , five claim 1 , six claim 1 , seven claim 1 , eight nine claim 1 , ten claim 1 , or all of the individual codons.3. The isolated recombinant nucleic acid of claim 2 , comprising at least 20 claim 2 , 30 claim 2 , 40 claim 2 , or 50 or more of the codons.4. The isolated recombinant nucleic acid of claim 1 , wherein the nucleotides are substituted such that a codon to produce Ala is GCG claim 1 , a codon to produce Cys is TGT claim 1 , a codon to produce Phe is TTT claim 1 , a codon to produce Pro is CCG claim 1 , a codon to produce Arg is CGT claim 1 , a codon to produce Ser is TCG claim 1 , ...

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Номер записи: 45
31-01-2019 дата публикации

Histidine Engineered Light Chain Antibodies and Genetically Modified Non-Human Animals for Generating the Same

Номер: US20190029237A1
Автор: MACDONALD Lynn, MCWHIRTER John, MURPHY Andrew J.
Принадлежит:

A genetically modified non-human animal is provided, wherein the non-human animal expresses an antibody repertoire capable of pH dependent binding to antigens upon immunization. A genetically modified non-human animal is provided that expresses human immunoglobulin light chain variable domains derived from a limited repertoire of human immunoglobulin light chain variable gene segments that comprise histidine modifications in their germline sequence. Methods of making non-human animals that express antibodies comprising histidine residues encoded by histidine codons introduced into immunoglobulin light chain nucleotide sequences are provided. 1. A genetically modified non-human animal comprising in its germline an immunoglobulin light chain locus comprising at least one human Vgene segment and at least one human Jgene segment operably linked to an immunoglobulin light chain constant region sequence ,{'sub': L', 'L, 'wherein each human Vgene segment comprises at least one histidine codon that is not encoded by the corresponding human germline Vgene segment, and'}{'sub': L', 'L, 'wherein the at least one human Vgene segment and the at least one human Jgene segment are capable of rearranging and encoding a human light chain variable domain of an antibody.'}222.-. (canceled)23. The genetically modified non-human animal of claim 1 , comprising in its germline an immunoglobulin light chain locus comprising no more than two human Vgene segments and one or more human Jgene segments operably linked to an immunoglobulin light chain constant region sequence claim 1 ,{'sub': L', 'L, 'wherein each of the no more than two human Vgene segments comprises at least one histidine codon that is not encoded by the corresponding human germline Vgene segment, and'}{'sub': L', 'L, 'wherein the human Vgene segments and Jgene segments are capable of rearranging and encoding a human light chain variable domain of an antibody.'}2448.-. (canceled)49. A method of generating an antibody light ...

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Номер записи: 46
01-02-2018 дата публикации

ANTIBODIES, VARIABLE DOMAINS & CHAINS TAILORED FOR HUMAN USE

Номер: US20180030121A1
Автор: Bradley Allan, England Nicholas, Friedrich Glenn, Lee E-Chiang, Strivens Mark
Принадлежит:

The invention relates to the provision of antibody therapeutics and prophylactics that are tailored specifically for human use. The present invention provides libraries, vertebrates and cells, such as transgenic mice or rats or transgenic mouse or rat cells. Furthermore, the invention relates to methods of using the vertebrates to isolate antibodies or nucleotide sequences encoding antibodies. Antibodies, heavy chains, polypeptides, nucleotide sequences, pharmaceutical compositions and uses are also provided by the invention. 1. A method for producing a heavy chain , VH domain or an antibody specific to a target antigen , wherein the heavy chain , VH domain or antibody comprises an HCDR3 at least 20 amino acids in length , the method comprisingproviding a non-human vertebrate, optionally a mouse or a rat, that has a genome comprising an immunoglobulin heavy chain locus comprising unrearranged human gene segment JH6*02, one or more VH gene segments and one or more D gene segments upstream of a constant region; wherein the gene segments in the heavy chain locus are operably linked to the constant region thereof so that the vertebrate is capable of producing an antibody heavy chain produced by recombination of the human JH6*02 with a human D segment and a human VH segment,wherein the JH6*02 gene segment comprises the following nucleotide sequence: ATTACTA CTACTACTAC GGTATGGACG TCTGGGGCCA AGGGACCACG GTCACCGTCT CCTCAG, andwherein the vertebrate has been immunised with the target antigen and produces an antibody heavy chain specific for the target antigen wherein the variable domain of the heavy chain is the product of recombination between a human VH, D and JH6*02 and wherein the HCDR3 length is at least 20 amino acids, andisolating from the non-human vertebrate the heavy chain, VH domain or an antibody specific to the target antigen or a cell producing the heavy chain, VH domain or antibody, wherein the heavy chain, VH domain or antibody comprises a HCDR3 that is ...

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Номер записи: 47
17-02-2022 дата публикации

AAV-IDUA VECTOR FOR TREATMENT OF MPS I-ASSOCIATED BLINDNESS

Номер: US20220047721A1
Автор: Hirsch Matthew Louis, Samulski Richard Jude
Принадлежит:

This invention relates to viral vectors for delivery of alpha-L-iduronidase to the cornea of a subject and methods of using the same for treatment and prevention of corneal clouding and blindness in a subject due to mucopolysaccharidosis I. 1. A recombinant nucleic acid comprising a sequence encoding human alpha-L-iduronidase (IDUA) , wherein the nucleotide sequence has been codon-optimized for expression in human cells.2. The recombinant nucleic acid of claim 1 , comprising a nucleotide sequence at least 90% identical to SEQ ID NO:1.3. The recombinant nucleic acid of claim 1 , comprising the nucleotide sequence of SEQ ID NO:1.4. An adeno-associated virus (NAV) vector genome comprising the nucleic acid of .5. The AAV vector genome of claim 4 , wherein the nucleic acid is operably linked to a constitutive promoter.6. A cell in vitro comprising the AAV vector genome of .7. The cell of claim 6 , wherein the vector genome is stably incorporated into the cell genome.8. An AAV particle comprising the AAV vector genome of .9. A method of producing a recombinant AAV particle comprising an AAV capsid claim 4 , the method comprising:{'claim-ref': {'@idref': 'CLM-00004', 'claim 4'}, 'providing a cell in vitro with AAV Cap and AAV Rep coding sequences, the AAV vector genome of , and helper functions for generating a productive AAV infection; and'}allowing assembly of the recombinant AAV particle comprising the AAV capsid and encapsidating the AAV vector genome.10. An AAV particle produced by the method of .11. The AAV particle of claim 8 , wherein the AAV particle is an AAV2 claim 8 , AAV8 claim 8 , or AAV9 particle.12. The AAV particle of claim 8 , wherein the AAV particle is a chimeric AAV8/AAV9 particle.13. A pharmaceutical formulation comprising the AAV particle of and a pharmaceutically acceptable carrier.14. A method of delivering claim 8 , IDUA to the cornea of a subject claim 8 , comprising administering to the cornea of the subject an effective amount of an AAV ...

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Номер записи: 48
17-02-2022 дата публикации

Transcriptional recording by crispr spacer acquisition from rna

Номер: US20220049232A1
Автор: Florian Schmidt, Randall Jeffrey Platt
Принадлежит: Eidgenoessische Technische Hochschule Zurich ETHZ

The present invention relates to a method for recording a transcriptome of a cell, the method comprising the steps of: providing a test cell comprising: a first transgene nucleic acid sequence encoding a fusion protein comprising a reverse transcriptase polypeptide and a Cas1 polypeptide and a second transgene nucleic acid sequence encoding a Cas2 polypeptide, wherein said first transgene nucleic acid sequence and said second transgene nucleic acid sequence are under transcriptional control of an inducible promoter sequence, and a third transgene nucleic acid sequence comprising a CRISPR direct repeat (DR) sequence; wherein said CRISPR direct repeat sequence is specifically recognizable by a RT-Cas1-Cas2 complex formed by the expression products of said first transgene nucleic acid sequence and said second transgene nucleic acid sequence, exposing said test cell to conditions under which expression of said first transgene nucleic acid sequence and said second transgene nucleic acid sequence is induced, wherein said RT-Cas1-Cas2 complex formed by expression products of said first transgene nucleic acid sequence and said second transgene nucleic acid sequence acquires protospacers from RNA molecules and integrates spacers into said third transgene nucleic acid sequence yielding a modified third transgene nucleic acid sequence, isolating said modified third transgene nucleic acid sequence from said test cell yielding an isolated third transgene nucleic acid sequence, and sequencing said isolated modified third transgene nucleic acid sequence.

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Номер записи: 49
31-01-2019 дата публикации

ANTI-NME ANTIBODY

Номер: US20190031778A1
Автор: Bamdad Cynthia, Smagghe Benoit
Принадлежит:

The present application discloses anti-NME antibodies and their use in treating or preventing diseases. 1. A method of treating or preventing cancer in a subject claim 79 , comprising administering to the subject the antibody of .25.-. (canceled)6. The method according to claim 1 , wherein the antibody inhibits binding between NME7 and its cognate binding partner.7. The method according to claim 6 , wherein the cognate binding partner is MUC1*.8. The method according to claim 6 , wherein the cognate binding partner is PSMGFR portion of the MUC1* extracellular domain.910.-. (canceled)11. The antibody according to claim 79 , wherein the peptide comprises a peptide claim 79 , which is highly homologous to claim 79 , or to which is added or subtracted up to 7 amino acid residues at the N-terminus or C-terminus.12. The antibody according to claim 11 , wherein the antibody is selected for its ability to bind to NME7-AB or NME7-X1 but not to NME1.13. The antibody according to claim 79 , wherein the antibody is polyclonal claim 79 , monoclonal claim 79 , bivalent claim 79 , monovalent claim 79 , bispecific claim 79 , an antibody fragment containing the variable region claim 79 , or an antibody mimic.14. The antibody according to claim 79 , wherein the antibody is human or humanized.15. The antibody according to claim 79 , wherein the antibody is a single chain scFv.1634.-. (canceled)35. A chimeric antigen receptor (CAR) claim 79 , for the treatment or prevention of cancer claim 79 , wherein the targeting extracellular portion of the chimeric antigen receptor comprises a portion of the antibody of .36. The chimeric antigen receptor according to claim 35 , wherein the portion of the antibody is a single chain scFv.37. The chimeric antigen receptor according to claim 35 , wherein the antibody is human or humanized.3878.-. (canceled)79. An antibody made against the peptide of (SEQ ID NOS:141 to 145). The present application relates to NME proteins, peptides derived from NME ...

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Номер записи: 50
31-01-2019 дата публикации

ANTI-CANCER PEPTIDE

Номер: US20190031779A1
Автор: Bamdad Cynthia, Smagghe Benoit
Принадлежит: MINERVA BIOTECHNOLOGIES CORPORATION

The present application discloses anti-NME antibodies and their use in treating or preventing diseases. 115.-. (canceled)16. A method of treating or preventing cancer in a subject claim 79 , comprising administering to the subject a peptide that is highly homologous or identical to regions of NME7-AB claim 79 , wherein the peptide is at least 80% homologous to the peptide of .1722.-. (canceled)23. The method according to claim 16 , wherein the peptide comprises a peptide claim 16 , which is highly homologous to claim 16 , or to which is added or subtracted up to 7 amino acid residues at the N-terminus or C-terminus.24. The method according to claim 16 , wherein the peptide is connected to another peptide via a spacer or linker.25. A chimeric antigen receptor (CAR) claim 79 , for the treatment or prevention of cancer wherein the targeting extracellular portion of the CAR comprises at least a peptide of .2631.-. (canceled)32. The chimeric antigen receptor according to claim 25 , wherein the peptide comprises a peptide claim 25 , which is highly homologous to claim 25 , or to which is added or subtracted up to 7 amino acid residues at the N-terminus or C-terminus.33. The chimeric antigen receptor according to claim 25 , wherein the peptide is connected to another peptide via a spacer or linker.34. A method of treating or preventing cancer or cancer metastasis claim 25 , comprising engineering the chimeric antigen receptor according to claim 25 , into an immune system cell and administering the cell to a subject in need thereof.3537.-. (canceled)38. A method of vaccinating a person against cancer or metastatic cancer comprising immunizing the person with a peptide fragment of .3944.-. (canceled)45. The method according to claim 38 , wherein the immunizing peptide comprises a peptide claim 38 , which is highly homologous to claim 38 , or to which is added or subtracted up to 7 amino acid residues at the N-terminus or C-terminus.46. The method according to claim 38 , ...

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Номер записи: 51
04-02-2021 дата публикации

MODULAR DNA ASSEMBLY SYSTEM

Номер: US20210032635A1
Автор: Nicholson Matthew Joseph, Parker Emily Jane, Van Dolleweerd Craig John
Принадлежит:

A modular and hierarchical DNA assembly platform for synthetic biology is described. This enabling technology, termed MIDAS (for Modular Idempotent DNA Assembly System), can precisely assemble multiple DNA fragments in a single reaction using a standardised assembly design. It can be used to build genes from libraries of sequence-verified, reusable parts and to assemble multiple genes in a single vector. We describe the design and use of MIDAS, and its application in the reconstruction of the metabolic pathway for production of paspaline, a key intermediate in the biosynthesis of a range of indole diterpenes—a class of economically important secondary metabolites produced by several species of filamentous fungi. 1. A vector set comprising at least two shuttle vectors , V1 and V2 , V1.1(+)=5′-A1 B1 M1 B2 C1 C2 A2-3′,', 'V1.2(−)=5′-A1 C1 C2 B1 M1 B2 A2-3′,', 'V1.3(+)=5′-A1 B2 M1 B1 C1 C2 A2-3′, and', 'V1.4(−)=5′-A1 C1 C2 B2 M1 B1 A2-3′,, 'wherein V1 is selected from the group consisting of'} V2.1(+)=5′-C1 B1 M1 B2 A1 M2 A2 C2-3′,', 'V2.2(−)=5′-C1 A1 M2 A2 B1 M1 B2 C2-3′,', 'V2.3(+)=5′-C1 B2 M1 B1 A1 M2 A2 C2-3′, and', 'V2.4(−)=5′-C1 A1 M2 A2 B2 M1 B1 C2-3′,, 'and V2 is selected from the group consisting of'}wherein M1 is a first marker, M2 is a second marker, and A1, A2, B1, B2, C1 and C2 are restriction enzyme recognition sites,wherein at least A1, A2, C1, and C2 are recognition sites for Type IIS restriction enzymes,wherein A1, B1 and C1 are all different recognition sites, andwherein A1=A2, C1=C2, and B1=B2 or B1≠B2.2. The vector set of comprising at least three claim 1 , at least four claim 1 , at least five claim 1 , at least six claim 1 , at least seven claim 1 , or eight different shuttle vectors.3. The vector set of wherein A1 claim 1 , A2 claim 1 , B1 claim 1 , B2 claim 1 , C1 and C2 comprise at least four claim 1 , preferably six claim 1 , Type IIS restriction sites.4. The vector set of further comprising a source vector or a destination vector or both claim ...

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Номер записи: 52
04-02-2021 дата публикации

PROMOTER AND USE THEREOF

Номер: US20210032636A1
Автор: Feng Aihua, Jia Yankai, Liao Guojuan, Qi Tianming, Sun Zhongping, Wu Xin, Xie Zhengli, Xue Gaoxu
Принадлежит:

An improved promoter and a use thereof. An improvement is to mutate a nucleic acid sequence between −35 region and −10 region in a promoter region into recognition sites for an endonuclease. The improvement is used for overcoming the problem that a transcription or translation product of foreign genes under a strong promoter might be toxic to a host and cannot be cloned and avoiding the phenomena of false positives and false negatives during blue-white screening. 1. An improved promoter , obtained by mutating a nucleic acid sequence between −35 region and −10 region in a promoter region into recognition sites for an endonuclease.2. The improved promoter of claim 1 , obtained by mutating a nucleic acid sequence between −35 region and −10 region in a promoter region of a β-galactosidase into the recognition sites for the endonuclease.3. The improved promoter of claim 2 , wherein the nucleic acid sequence between −35 region and −10 region in the promoter region of the β-galactosidase is shown by SEQ ID NO.1-2.4. The improved promoter of claim 2 , wherein the endonuclease is any one or a combination of at least two of EcoRV claim 2 , AleI claim 2 , BamHI claim 2 , XhoI and PmlI.5. The improved promoter of claim 2 , wherein a nucleic acid sequence between −35 region and −10 region of the improved promoter is shown by SEQ ID NO.3-14.6. A vector claim 1 , comprising the improved promoter of .7. The vector of claim 6 , further comprising: a gene of interest claim 6 , wherein the gene of interest is operably ligated between the recognition sites for the endonuclease of the improved promoter.8. The vector of claim 7 , wherein the vector is a cloning vector and/or an expression vector claim 7 , preferably the cloning vector.9. A host cell claim 6 , comprising the vector of .10Escherichia coli.. The host cell of claim 9 , wherein the host cell is11Escherichia coli. The host cell of claim 10 , wherein a C-terminal ω-fragment of a β-galactosidase of the is only encoded.12. A ...

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Номер записи: 53
04-02-2021 дата публикации

EXPRESSION OF PHYTASE IN ASPERGILLUS NIGER

Номер: US20210032637A1
Автор: BAI Aixi, BIAN Furong, Li Feng, Sun Yan, Xu Hong, ZHU Jidong
Принадлежит:

Disclosed herein is a method for expressing phytase in a filamentous fungus by using an optimized phytase gene having a nucleotide sequence as shown in SEQ ID NO. 7 and a signal peptide having a nucleotide sequence as shown in SEQ ID NO. 12. 1Escherichia coliAspergillus oryzae. A signal peptide for enhancing the secretory expression of phytase or a mutant thereof in a filamentous fungus , wherein the signal peptide is derived from TAKA amylase and has an amino acid sequence as shown in SEQ ID NO. 13.2Escherichia coli. The signal peptide for enhancing the secretory expression of phytase or a mutant thereof in a filamentous fungus according to claim 1 , having a nucleotide sequence as shown in SEQ ID NO. 12.3Escherichia coliAspergillus niger.. The signal peptide for enhancing the secretory expression of phytase or a mutant thereof in a filamentous fungus according to claim 1 , wherein the filamentous fungus is selected from4Escherichia coliEscherichia coli. The signal peptide for enhancing the secretory expression of phytase or a mutant thereof in a filamentous fungus according to claim 3 , wherein the phytase has an amino acid sequence as shown in SEQ ID NO. 4.5Escherichia coliEscherichia coli. The signal peptide for enhancing the secretory expression of phytase or a mutant thereof in a filamentous fungus according to claim 3 , wherein the mutant of the phytase has an amino acid sequence as shown in SEQ ID NO. 15 or SEQ ID NO. 17.6Escherichia coli. A codon-optimized gene encoding phytase or a mutant thereof claim 3 , which has a nucleotide sequence as shown in SEQ ID NO. 7; or has a nucleotide sequence that is at least 95% claim 3 , 96% claim 3 , 97% claim 3 , 98% or 99% homologous to the nucleotide sequence as shown in SEQ ID NO. 7 claim 3 , and encodes a protein having the phytase activity.7Escherichia coli. A codon-optimized DNA sequence encoding phytase or a mutant mature peptide thereof claim 3 , which has a nucleotide sequence as shown in SEQ ID NO. 8; or has a ...

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Номер записи: 54
24-02-2022 дата публикации

AAV-EPO FOR TREATING COMPANION ANIMALS

Номер: US20220056090A1
Автор: Hinderer Christian, Wilson James M., Wilson Matthew
Принадлежит:

Compositions and methods are provided for treating companion animals are provided. An adeno-associated viral vector is provided which includes a nucleic acid molecule comprising a sequence encoding erythropoietin (EPO). In desired embodiments, the subject is a cat or dog. 1. A recombinant adeno-associated virus (rAAV) comprising an AAV capsid having packaged therein a vector genome , wherein said vector genome comprises nucleic acid sequences encoding feline or canine erythropoietin (EPO) , inverted terminal repeat sequences and expression control sequences that direct expression of the EPO in a host cell.2. A method for treating chronic kidney disease , said method comprising administering to a subject in need thereof a pharmaceutical composition comprising a pharmaceutically acceptable carrier and a recombinant adeno-associated virus (rAAV) comprising an AAV capsid having packaged therein a vector genome , wherein said vector genome comprises a nucleic acid sequence encoding feline or canine erythropoietin (EPO) , inverted terminal repeat sequences , and expression control sequences that direct expression of the EPO in a host cell.3. The method according to claim 2 , wherein the nucleotide sequence encoding the feline EPO comprises nucleotides 79 to 576 of the nucleotide sequence of SEQ ID NO: 8 and encodes at least amino acids 27 to 192 of the amino acid sequence of SEQ ID NO: 4.4. The method according to claim 2 , wherein the encoded feline EPO comprises the amino acids 27 to 192 of SEQ ID NO: 4 in combination with a heterologous leader sequence.5. The rAAV of claim 2 , wherein the EPO nucleotide sequence encodes amino acids 1 to 192 of SEQ ID NO: 4.6. The rAAV of claim 5 , wherein the nucleotide sequence encoding the feline EPO comprises SEQ ID NO: 8.7. The method according to claim 2 , wherein the nucleotide sequence encoding the canine EPO comprises nucleotides 121 to 618 of the nucleotide sequence of SEQ ID NO: 7 and encodes at least amino acids 41 to 206 of ...

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Номер записи: 55
12-02-2015 дата публикации

Treatment methods using adenovirus

Номер: US20150044280A1
Автор: Andrea Leubitz, Erez Feige, Eyal Breitbart, Richard PENSON
Принадлежит: Vascular Biogenics Ltd

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.

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Номер записи: 56
07-02-2019 дата публикации

COMMON LIGHT CHAIN MOUSE

Номер: US20190040123A1
Автор: BUCKLER David R., MACDONALD Lynn, MCWHIRTER John, MURPHY Andrew J., STEVENS Sean
Принадлежит:

A genetically modified mouse is provided, wherein the mouse is incapable of rearranging and expressing an endogenous mouse immunoglobulin light chain variable sequence, wherein the mouse expresses only one or two human light chain variable domains encoded by human immunoglobulin sequences operably linked to the mouse kappa (κ) constant gene at the endogenous mouse κ locus, wherein the mouse expresses a reverse chimeric antibody having a light chain variable domain derived from one of only two human light chain variable region gene segments and a mouse κ constant domain, and a human heavy chain variable domain and a mouse heavy chain constant domain, from an endogenous mouse heavy chain locus. Bispecific epitope-binding proteins that are fully human are provided, comprising two different heavy chains that associate with an identical light chain that comprises a variable domain derived from one of two different human light chain variable region gene segments. 121.-. (canceled)22. A genetically modified mouse that is homozygous or heterozygous in its germline genome for a single rearranged human immunoglobulin kappa light chain variable region sequence comprising a human Vκ gene segment and a human Jκ gene segment ,wherein all immunoglobulin kappa light chains expressed by B cells of the genetically modified mouse comprise immunoglobulin kappa light chain variable domains expressed from the single rearranged human immunoglobulin kappa light chain variable region sequence or a somatically hypermutated version thereof, and wherein the genetically engineered mouse lacks endogenous immunoglobulin Vκ and/or Jκ gene segments that are capable of rearranging to form an endogenous immunoglobulin light chain variable region sequence.23. The genetically modified mouse of claim 22 , wherein the single rearranged human immunoglobulin kappa light chain variable region sequence is a Vκ1-39/Jκ sequence.24. The genetically modified mouse of claim 23 , wherein the single rearranged ...

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Номер записи: 57
15-02-2018 дата публикации

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

Номер: US20180043034A1
Автор: Farkas Michael H., Pierce Eric A., Sousa Maria E.
Принадлежит:

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 ...

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Номер записи: 58
03-03-2022 дата публикации

Codon optimized rep1 genes and uses thereof

Номер: US20220062438A1
Автор: David H. Kirn, Melissa A. KOTTERMAN, Peter Francis
Принадлежит: 4D Molecular Therapeutics Inc

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.

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Номер записи: 59
25-02-2021 дата публикации

Genetically modified t cell receptor mice

Номер: US20210051930A1
Автор: Andrew J. Murphy, Cagan Gurer, John Mcwhirter, Karolina MEAGHER, Lynn Macdonald, Naxin Tu, Sean Stevens, Vera VORONINA
Принадлежит: Regeneron Pharmaceuticals Inc

The invention provides a genetically modified non-human animal that comprises in its genome unrearranged T cell receptor variable gene loci, as well as embryos, cells, and tissues comprising the same. Also provided are constructs for making said genetically modified non-human animal and methods of making the same. Various methods of using the genetically modified non-human animal are also provided.

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Номер записи: 60
14-02-2019 дата публикации

Methods and materials for producing immune responses against polypeptides involved in antibiotic resistance

Номер: US20190046629A1
Автор: Michael A. Barry
Принадлежит: Mayo Foundation for Medical Education and Research

This document relates to methods and materials for producing immune responses against polypeptides involved in antibiotic resistance. For example, vaccines against polypeptides involved in antibiotic resistance as well as methods for vaccinating mammals against polypeptides involved in antibiotic resistance are provided.

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Номер записи: 61
25-02-2016 дата публикации

Recombinant polynucleotide and a transgenic flammulina velutipes carrying the same

Номер: US20160051664A1
Автор: Ying-Tzu Lyu
Принадлежит: MycoMagic Biotechnology Co Ltd

The invention provides a recombinant polynucleotide comprising a truncated glyceraldehyde-3-phosphate dehydrogenase (gpd) promoter and a modified HBV S protein gene and a transgenic Flammulina velutipes carrying the recombinant polynucleotide. The invention surprisingly found that after administering the transgenic Flammulina velutipes to a subject, the subject can successfully generate an antibody against HBV. Therefore, the transgenic Flammulina velutipes can be used as a vaccine against HBV.

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Номер записи: 62
23-02-2017 дата публикации

Compositions useful in treatment of ornithine transcarbamylase (otc) deficiency

Номер: US20170051259A1
Автор: James M Wilson, Lili Wang
Принадлежит: University of Pennsylvania Penn

Viral vectors comprising engineered hOTC DNA and RNA sequences are provided which when delivered to a subject in need thereof are useful for treating hyperammonemia, ornithine transcarbamylase transcarbamylase deficiency and symptoms associated therewith. Also provided are methods of using hOTC for treatment of liver fibrosis cirrhosis in OTCD patients by administering hOTC.

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Номер записи: 63
26-02-2015 дата публикации

Methods for making fully human bispecific antibodies using a common light chain

Номер: US20150059009A1
Автор: Andrew J. Murphy, John Mcwhirter, Lynn Macdonald, Samuel Davis, Sean Stevens
Принадлежит: Regeneron Pharmaceuticals Inc

A genetically modified mouse is provided, wherein the mouse expresses an immunoglobulin light chain repertoire characterized by a limited number of light chain variable domains. Mice are provided that express just one or a few immunoglobulin light chain variable domains from a limited repertoire in their germline. Methods for making bispecific antibodies having universal light chains using mice as described herein, including human light chain variable regions, are provided. Methods for making human variable regions suitable for use in multispecific binding proteins, e.g., bispecific antibodies, and host cells are provided. Bispecific antibodies capable of binding first and second antigens are provided, wherein the first and second antigens are separate epitopes of a single protein or separate epitopes on two different proteins are provided.

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Номер записи: 64
21-02-2019 дата публикации

TREATMENT METHODS USING ADENOVIRUS

Номер: US20190054130A1
Автор: Breitbart Eyal, FEIGE EREZ, LEUBITZ Andrea, Penson Richard
Принадлежит: Vascular Biogenics Ltd.

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 ...

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Номер записи: 65
10-03-2022 дата публикации

ADENOVIRAL-BASED BIOLOGICAL DELIVERY AND EXPRESSION SYSTEM FOR USE IN THE TREATMENT OF OSTEOARTHRITIS

Номер: US20220073948A1
Автор: Guse Kilian, Lee Brendan, Ruan Zhechao
Принадлежит:

The invention relates to an adenoviral-based biological delivery and expression system for use in the treatment or prevention of osteoathritis in human or mammalian joints by long-term inducible gene expression of human or mammalian interleukin-1 receptor antagonist (II-1 Ra) in synovial cells, comprising a helper-dependent adenoviral vector containing a nucleic acid sequence encoding for human or mammalian interleukin-1 receptor antagonist (II-1 Ra), left and right inverted terminal repeats (L ITR and R ITR), the adenoviral packaging signal and non-viral, non-coding stuffer nucleic acid sequences, wherein the expression of the human or mammalian interleukin-1 receptor antagonist (II-1 Ra) gene within synovial cells is regulated by an inflammation-inducible promoter. 1. An adenoviral-based biological delivery and expression system for use in the treatment or prevention of osteoathritis in human or mammalian joints by long-term inducible gene expression of human or mammalian interleukin-1 receptor antagonist (II-1Ra) in synovial cells , comprising a helper-dependent adenoviral vector containing a nucleic acid sequence encoding for human or mammalian interleukin-1 receptor antagonist (II-1Ra) , left and right inverted terminal repeats (L ITR and R ITR) , the adenoviral packaging signal and non-viral , non-coding stuffer nucleic acid sequences , wherein the expression of the human or mammalian interleukin-1 receptor antagonist (II-1Ra) gene within synovial cells is regulated by an inflammation-inducible promoter , which is located upstream of the reading frame of the nucleic acid sequence encoding for human or mammalian interleukin-1 receptor antagonist (II-1Ra) and which is specifically activated by increased levels of immune stimulatory substances.2. The adenoviral-based biological delivery and expression system according to claim 1 , wherein the inflammation-inducible promoter is selected from the group consisting of NF-κB promoter claim 1 , interleukin 6 (II-6) ...

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Номер записи: 66
04-03-2021 дата публикации

Methods and compositions for treatment of ocular disorders and blinding diseases

Номер: US20210060176A1
Автор: Jean Bennett, Jeannette BENNICELLI, JUNWEI Sun
Принадлежит: University of Pennsylvania Penn

Codon optimized nucleic acid sequences for the long form and short form of RdCVF are provided, as well as recombinant viral vectors, such as AAV, expression cassettes, proviral plasmids or other plasmids containing the codon optimized sequences. Recombinant vectors are provided that express the codon optimized RdCVFL and RdCVF individually, or express two copies of a codon optimized RdCVF or RdCVFL nucleic acid sequence, or both RdCVFL and RdCVF in a single vector or virus. Compositions containing these codon optimized sequences are useful in methods for treating, retarding or halting certain blinding diseases resulting from the absence or inappropriate expression of RdCVF and RdCVFL.

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Номер записи: 67
21-02-2019 дата публикации

Vector Comprising Multiple Homologous Nucleotide Sequences

Номер: US20190055582A1
Автор: Steigerwald Robin
Принадлежит: BAVARIAN NORDIC A/S

The invention relates to vectors comprising two or more homologous nucleotide sequences and methods for generating them. The invention concerns substituting bases in the homologous nucleotide sequences with different bases that do not alter the encoded amino acid sequence. The invention allows for the reduction of intramolecular recombination between homologous nucleotide sequences, in particular in mammalian cells. The invention further relates to nucleotide sequences containing substituted bases. 119.-. (canceled)20. A recombinant modified vaccinia Ankara (MVA) virus vector that stably encodes homologous sequences , the vector comprising:first and second nucleotide sequences of at least 1500 nucleotides each, each coding for at least 500 amino acids, wherein at least 150 continuous amino acids encoded by each of the two nucleotide sequences have at least 75% amino acid identity;wherein at least one of the first and second nucleotides has at least 400 substituted nucleotides and wherein the substituted nucleotides do not alter the identical amino acids encoded by said two nucleotide sequences; andwherein the first and second nucleotide sequences differ by at least 400 nucleotides; andwherein the first and second nucleotides share stretches of identity of no more than 9 contiguous nucleotides; and wherein the first and second nucleotide sequences each encode a RSV protein.21. The recombinant MVA virus vector of claim 20 , wherein first and second nucleotide sequences encode a full-length RSV-F protein and a truncated RSV-F protein.22. The recombinant MVA virus vector of claim 20 , wherein the first and second nucleotide sequences encode the amino acid sequences of SEQ ID NO:3 and SEQ ID NO:4 claim 20 , respectively.23. The recombinant MVA virus vector of claim 22 , wherein the first and second nucleotide sequences comprise the nucleotide sequences of SEQ ID NO: 1 and SEQ ID NO:2 claim 22 , respectively. This application is a continuation application of U.S. ...

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Номер записи: 68
01-03-2018 дата публикации

DNA PLASMID, CODING HNP-1, OR HNP-2, OR HNP-3, BACTERIAL PRODUCER, ANALGESIC AGENT (VARIANTS)

Номер: US20180057837A1
Автор: DUKHOVLINOV Ilya Vladimirovich, ORLOV Anton Iosifovich
Принадлежит:

The invention (embodiments) relates to the field of medicine, pharmacology, biotechnology, molecular biology, genetic engineering and can be used for analgesia. A plasmid DNA is given for transient expression in mammalian cells, represented by a skeleton that contains prokaryotic and eukaryotic elements and a polynucleotide encoding human alpha defensin HNP-1 or HNP-2 or HNP-3 (embodiments). A producer of such plasmid DNA is also given based on a bacterial cell (embodiments) and an analgesic agent for use in mammals, in particular, human, based on it (embodiments). The technical result from the use of embodiments of the invention is expressed in expanding the range of analgesic products, in an increase of the duration of analgesia, in an increase of the safety of analgesia, in simplification and cheapening of the production of the analgesic. 1. A plasmid DNA for transient expression in mammalian cells , represented by a skeleton that contains prokaryotic elements , an origin of replication and a reporter gene , and eukaryotic elements , a strong promoter , a leader sequence of mRNA , and also regulatory sequences for the mentioned elements , at least one site for cloning of a gene of interest and at least one site for annealing of at least one primer for analysis of the composition of the plasmid DNA , and a polynucleotide represented by a secretory sequence , a fragment encoding human alpha defensin HNP-1 or HNP-2 or HNP-3 , codon-optimized for expression in mammalian cells , and a termination sequence.2. Plasmid DNA according to claim 1 , characterized by the fact that preproprotein is represented by SEQ ID NO:1 or SEQ ID NO:2 or SEQ ID NO:7 or SEQ ID NO:8.3. Plasmid DNA according to claim 1 , characterized in that the polynucleotide is represented by SEQ ID NO:3 or SEQ ID NO:4 or SEQ ID NO:5 or SEQ ID NO:6 or SEQ ID NO:9 or SEQ ID NO: 10 or SEQ ID NO:11 or SEQ ID NO: 12.4. A producer of the plasmid DNA according to claim 1 , based on a bacterial cell.5. An ...

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Номер записи: 69
01-03-2018 дата публикации

VECTORS AND METHODS FOR TARGETED INTEGRATION IN LOCI COMPRISING CONSTITUTIVELY EXPRESSED GENES

Номер: US20180057842A1
Автор: ELEFANTY Andrew, Elliott David, LABONNE Tatiana, STANLEY Ed
Принадлежит:

The invention relates to a vector comprising: a 5′ nucleic acid that is homologous to a genomic sequence 5′ of a stop codon of a constitutively expressed gene; an exogenous nucleic acid; a 3′ nucleic acid that is homologous to a genomic sequence 3′ of the stop codon of the constitutively expressed gene; a translation interruption-reinitiation signal operably linked to the 5′ nucleic acid and the exogenous Targeting nucleic acid, wherein the translation interruption-reinitiation signal is capable of replacing the stop codon of the constitutively expressed gene. 1. A vector comprising:a 5′ nucleic add that is homologous to a genomic sequence 5′ of a genomic stop codon of a constitutively expressed gene;an exogenous nucleic acid;a 3′ nucleic acid that is homologous to a genomic sequence 3′ of the genomic stop codon of the constitutively expressed gene;a translation interruption-reinitiation signal operably linked to the 5′ nucleic acid and the exogenous nucleic acid,wherein the translation interruption-reinitiation signal is capable of replacing the stop codon of the constitutively expressed gene.2. The vector of claim 1 , wherein the constitutively expressed gene comprises GAPDH claim 1 , ACTB claim 1 , HSP90 claim 1 , B2M claim 1 , HPRTI claim 1 , RPLP1 claim 1 , GUSB claim 1 , LDHA claim 1 , NONO claim 1 , PGK1 claim 1 , PPIH claim 1 , RPLPO claim 1 , or TFRC.3. The vector of claim 1 , wherein the translation interruption-reinitiation signal encodes a 2A peptide.4. The vector of claim 3 , wherein the translation inten p io reinitiation signal comprises SEQ ID NO: 1.5. The vector of claim 1 , wherein the 5′ nucleic add comprises about 25 to 100 bases claim 1 , about 200 to 1000 bases claim 1 , about 1 kb to 5 kb claim 1 , or about 3.5 kb.16. The vector of claim 5 , wherein the 5′ nucleic acid comprises SEQ ID NO: 2.7. The vector of claim 1 , wherein the 3′ nucleic acid comprises about 25 to 100 bases claim 1 , about 200 to 1000 bases claim 1 , about 1 kb to 5 kb ...

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Номер записи: 70
02-03-2017 дата публикации

Engineered CRISPR-Cas9 Nucleases

Номер: US20170058271A1
Автор: Benjamin Kleinstiver, J. Keith Joung, Vikram PATTANAYAK
Принадлежит: General Hospital Corp

Engineered CRISPR-Cas9 nucleases with improved specificity and their use in genomic engineering, epigenotnic engineering, genome targeting, and genome editing.

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Номер записи: 71
04-03-2021 дата публикации

GENETICALLY MODIFIED RECOMBINANT VACCINIA ANKARA (RMVA) VACCINES OF IMPROVED STABILITY AND METHODS OF PREPARATION THEREOF

Номер: US20210062221A1
Автор: CONTRERAS Heidi, DIAMOND Don J., WUSSOW Felix
Принадлежит: CITY OF HOPE

A vaccine composition comprising an immunologically effective amount of recombinant modified vaccinia Ankara (rMVA) virus comprising IE1, IE2 and pp65 or antigenic fragments thereof, which is genetically stable after at least 10 passages. A method of improving the stability of such rMVA upon passage by including one or more of the modifications: (1) inserting one or more nucleic acid sequences encoding the CMV antigens or antigenic fragments thereof into one or more insertion sites including but not limited to 044L/045L, IGR3, G1L/I8R, and Del3 but not including Del2; (2) codon optimizing the nucleic acid sequences encoding the CMV antigens by removing consecutive cytosines or guanines; and (3) introducing one or more mutations in the amino acid sequences of the CMV antigens. 1. An expression system for co-expressing two or more cytomegalovirus (CMV) antigens comprising a genetically modified recombinant Vaccinia Ankara (rMVA) vector inserted with two or more nucleic acid sequences encoding two or more CMV antigens or antigenic portions thereof , wherein the CMV antigens or antigenic fragments thereof include IE1 exon 4 (IE1/e4) , IE2 exon 5 (IE2/e5) , IEfusion , and pp65 , and wherein the two or more nucleic acid sequences are inserted in one or more insertion sites selected from 044L/045L , IGR3 , G1L/I8R , and Del3.2. The expression system of claim 1 , wherein the two or more nucleic acid sequences are operably linked to and under the control of a single promoter.3. The expression system of claim 2 , wherein the promoter is an mH5 promoter.4. The expression system of any one of - claim 2 , wherein one or more nucleic acid sequences are codon optimized to remove consecutive cytosines or guanines while expressing the same amino acids.5. The expression system of any one of - claim 2 , wherein the amino acid sequences of the CMV antigens comprise one or more mutations to improve the genetic stability of the rMVA upon viral passaging.6. The expression system of any ...

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Номер записи: 72
20-02-2020 дата публикации

Compositions Useful in Treatment of Spinal Muscular Atrophy

Номер: US20200056205A1
Автор: Hinderer Christian, Nathan Katz, Wang Qiang, Wilson James M.
Принадлежит:

A rAAV vector is described herein which has an AAVhu68 capsid and at least one expression cassette in the capsid. The at least one expression cassette comprises nucleic acid sequences encoding a functional SMN protein and expression control sequences that direct expression of the SMN sequences in a host cell. Also provided are compositions containing this rAAVhu68.SMN vector and methods of using same for spinal muscular atrophy in a patient. 1. A recombinant adeno-associated viral (rAAV) vector comprising an AAVhu68 capsid and at least one expression cassette , wherein the at least one expression cassette comprises nucleic acid sequences encoding a functional SMN protein and expression control sequences that direct expression of the SMN sequences in a host cell , wherein the rAAV has an AAVhu68 capsid comprising:a heterogenous population of AAVhu68 vp1 proteins selected from: vp1 proteins produced by expression from a nucleic acid sequence which encodes the predicted amino acid sequence of 1 to 736 of SEQ ID NO:8, vp1 proteins produced from SEQ ID NO: 7, or vp1 proteins produced from a nucleic acid sequence at least 70% identical to SEQ ID NO:7 which encodes the predicted amino acid sequence of 1 to 736 of SEQ ID NO:8,a heterogenous population of AAVhu68 vp2 proteins selected from: vp2 proteins produced by expression from a nucleic acid sequence which encodes the predicted amino acid sequence of at least about amino acids 138 to 736 of SEQ ID NO:8, vp2 proteins produced from a sequence comprising at least nucleotides 412 to 2211 of SEQ ID NO:7, or vp2 proteins produced from a nucleic acid sequence at least 70% identical to at least nucleotides 412 to 2211 of SEQ ID NO:7 which encodes the predicted amino acid sequence of at least about amino acids 138 to 736 of SEQ ID NO:8, anda heterogenous population of AAVhu68 vp3 proteins selected from: vp3 produced by expression from a nucleic acid sequence which encodes the predicted amino acid sequence of at least about amino ...

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Номер записи: 73
27-02-2020 дата публикации

GENE THERAPY FOR OCULAR DISORDERS

Номер: US20200061209A1
Автор: Bennett Jean, Bennicelli Jeannette, Sun Junwei
Принадлежит:

Compositions and methods are provided for treating ocular disorders in a subject are provided. In one aspect, an adeno-associated viral vector is provided which includes a nucleic acid molecule comprising a sequence encoding CNGA3. In another aspect, an adeno-associated viral vector is provided which includes a nucleic acid molecule comprising a sequence encoding CNGB3. In another aspect, an adeno-associated viral vector is provided which includes a nucleic acid molecule comprising a sequence encoding REP-1. In desired embodiments, the subject is human, cat, dog, sheep, or non-human primate. 13-. (canceled)4. An adeno-associated virus (AAV) vector comprising an AAV capsid and a nucleic acid sequence comprising AAV inverted terminal repeat sequences and a nucleic acid sequence encoding human Rab Escort Protein-1 (REP-1) , and expression control sequences that direct expression of the REP-1 in a host cell.5. (canceled)6. The viral vector of claim 4 , wherein the REP-1 sequence encodes the protein sequence of SEQ ID NO: 2.78-. (canceled)9. The viral vector of claim 4 , wherein the REP-1 sequence comprises SEQ ID NO: 1.1012-. (canceled)13. An adeno-associated virus (AAV) vector comprising an AAV capsid and a nucleic acid sequence comprising AAV inverted terminal repeat sequences and a nucleic acid sequence encoding human cyclic nucleotide gated channel alpha 3 (CNGA3) claim 4 , and expression control sequences that direct expression of the CNGA3 in a host cell.14. (canceled)15. The viral vector of claim 13 , wherein the CNGA3 sequence encodes the protein sequence of SEQ ID NO: 10.1617-. (canceled)18. The viral vector of claim 13 , wherein CNGA3 sequence comprises SEQ ID NO: 9 or SEQ ID NO: 11.19. The viral vector of any of claim 4 , wherein the expression control sequences comprise a chicken β-actin (CBA) promoter with cytomegalovirus (CMV) enhancer elements.20. (canceled)21. The viral vector of claim 9 , wherein the promoter is a rhodopsin kinase promoter.22. The viral ...

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Номер записи: 74
28-02-2019 дата публикации

HIGHLY EFFICIENT INFLUENZA MATRIX (M1) PROTEINS

Номер: US20190062713A1
Автор: KORT Thomas, MASSARE Michael J., NATHAN Margaret, PUSHKO Peter M., ROBINSON Robin, SMITH Gale, Wu Yingyun
Принадлежит:

This invention discloses a method of increasing production of virus-like particles comprising expressing an avian influenza matrix protein. The invention also comprises methods of making and using said VLPs. 1. A virus-like particle (VLP) comprising influenza proteins , wherein the influenza proteins consist of an influenza M1 protein and an influenza hemagglutinin (HA) protein , wherein said M1 protein comprises the amino acid residues YKKL at the positions corresponding to positions 100-103 of SEQ ID NO: 49.2. The VLP of claim 1 , wherein said influenza M1 protein is derived from an avian influenza virus strain.3. The VLP of claim 2 , wherein said avian influenza virus strain is A/Indonesia/5/05.4. The VLP of claim 1 , wherein the HA protein is from a different influenza strain than the M1 protein.5. The VLP of claim 4 , wherein said HA protein is derived from an avian influenza virus.6. The VLP of claim 5 , wherein said avian influenza virus is H5N1.7. The VLP of claim 5 , wherein said avian influenza virus is H9N2.8. The VLP of claim 4 , wherein said HA protein is derived from a seasonal influenza virus.9. The VLP of claim 8 , wherein said seasonal influenza virus is a type A influenza virus.10. The VLP of claim 8 , wherein said seasonal influenza virus is a type B influenza virus.11. The VLP of claim 4 , wherein said HA protein exhibits hemagglutinin activity.12. The VLP of claim 4 , wherein said HA protein is a chimeric protein.13. The VLP of claim 12 , wherein said chimeric proteins comprise external domains of non-avian influenza HA protein sequences fused to the transmembrane and/or cytoplasmic terminal domains of the avian HA.14. The VLP of claim 13 , wherein said non-avian influenza HA is from influenza strain A/Wisconsin/67/2005 and said avian influenza HA is from influenza strain A/Indonesia/5/05.15. A method of increasing the efficiency of VLP production comprising expressing an influenza M1 protein and an influenza hemagglutinin (HA) protein in an ...

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Номер записи: 75
28-02-2019 дата публикации

Vectors and host cells comprising a modified SV40 promoter for protein expression

Номер: US20190062797A1
Автор: HARTMAN Taymar E., HE YIMIN, Tso J. Yun
Принадлежит:

The present disclosure is directed to expression vectors, comprising a weakened SV40 promoter, and recombinant mammalian cells capable of producing high levels of a polypeptide of interest, methods of generating and using such recombinant mammalian cells. 1. A vector comprising a dESV40 promoter operably linked to a polypeptide coding sequence.2. The vector of claim 1 , in which the dESV40 promoter corresponds in sequence to SEQ ID NO: 1.3. A vector comprising:(i) an expression cassette, where the expression cassette comprises a first polynucleotide sequence encoding a first polypeptide of interest operably linked to a first expression promoter capable of effecting expression of the first polypeptide in a mammalian cell; and(ii) a polynucleotide sequence encoding a selectable marker operably linked to a dESV40 promoter.4. The vector of claim 3 , in which the expression cassette further comprises a second polynucleotide sequence encoding a second polypeptide of interest operably linked to a second expression promoter capable of effecting expression of the second polypeptide in a mammalian cell.5. The vector of claim 4 , in which the first and second expression promoters are the same.6. The vector of claim 4 , claim 4 , or claim 4 , in which the first and the second expression promoters are selected from the group consisting of: Rous sarcoma virus (RSV) promoter claim 4 , cytomegalovirus immediate early (CMV IE) promoter claim 4 , cytomegalovirus major intermediate-early (MIE) promoter claim 4 , simian virus 40 (SV40) early promoter claim 4 , Adenovirus major late promoter (AML) claim 4 , mouse mammary tumor virus LTR promoter claim 4 , Herpes thymidine kinase promoter claim 4 , human β-actin (ACTB) promoter claim 4 , elongation factor-1α (EF1α) promoter claim 4 , the phosphoglycerate kinase (PGK) promoter claim 4 , the ubiquitinC (UbC) promoter claim 4 , and murine metallotheionin promoter.7. The vector of claim 4 , claim 4 , or claim 4 , in which the selectable ...

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Номер записи: 76
09-03-2017 дата публикации

Sequential administration of a replication defective adenovirus vector in vaccination protocols

Номер: US20170065693A1
Автор: Balint Joseph P., Gayle Richard B., Jones Frank R.
Принадлежит:

Methods for generating immune responses using adenovirus vectors that allow multiple vaccinations with the same adenovirus vector and vaccinations in individuals with preexisting immunity to adenovirus are provided. 136-. (canceled)37. A composition comprising a replication defective adenovirus vector comprising:a deletion in the E2b region; anda nucleic acid sequence encoding Her2/neu.38. The composition of claim 37 , further comprising a replication defective adenovirus vector comprising a nucleic acid sequence encoding a tumor protein claim 37 , a viral coat protein claim 37 , a bacterial surface protein claim 37 , or a combination thereof.39. The composition of claim 37 , wherein the replication defective adenovirus vector further comprises a deletion in the E1 region claim 37 , a deletion in the E3 region claim 37 , a deletion in the E4 region claim 37 , or a combination thereof.40. The composition of claim 37 , wherein the replication defective adenovirus vector is not a gutted vector.41. The composition of claim 37 , wherein the composition comprises the replication defective adenovirus vector at a concentration of at least 10virus particles/ml.42. The composition of claim 37 , wherein the composition comprises the replication defective adenovirus vector at a concentration of at least 10virus particles/ml.43. The composition of claim 37 , wherein the composition comprises the replication defective adenovirus vector at a concentration of at least 10virus particles/ml.44. The composition of claim 37 , wherein the composition comprises the replication defective adenovirus vector at a concentration of at least 10virus particles/ml.45. The composition of claim 37 , wherein the composition comprises the replication defective adenovirus vector at a concentration of at least 10virus particles/ml.46. The composition of claim 37 , wherein the composition comprises the replication defective adenovirus vector at a concentration of at least 10virus particles/ml.47. The ...

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Номер записи: 77
09-03-2017 дата публикации

METHODS AND COMPOSITIONS FOR PRODUCING AN ADENOVIRUS VECTOR FOR USE WITH MULTIPLE VACCINATIONS

Номер: US20170065706A1
Автор: Balint Joseph P., Gayle Richard B., Jones Frank R.
Принадлежит:

Methods for generating immune responses using adenovirus vectors that allow multiple vaccinations with the same adenovirus vector and vaccinations in individuals with preexisting immunity to adenovirus are provided. 136-. (canceled)37. A composition comprising a replication defective adenovirus vector comprising:a deletion in the E2b region; anda nucleic acid sequence encoding HPV E6, HPV E7 or a combination thereof.38. The composition of claim 37 , further comprising a replication defective adenovirus vector comprising a nucleic acid sequence encoding a tumor protein claim 37 , a viral coat protein claim 37 , a bacterial surface protein claim 37 , or a combination thereof.39. The composition of claim 37 , wherein the replication defective adenovirus vector further comprises a deletion in the E1 region claim 37 , a deletion in the E3 region claim 37 , a deletion in the E4 region claim 37 , or a combination thereof.40. The composition of claim 37 , wherein the replication defective adenovirus vector is not a gutted vector.41. The composition of claim 37 , wherein the composition comprises the replication defective adenovirus vector at a concentration of at least 10virus particles/ml.42. The composition of claim 37 , wherein the composition comprises the replication defective adenovirus vector at a concentration of at least 10virus particles/ml.43. The composition of claim 37 , wherein the composition comprises the replication defective adenovirus vector at a concentration of at least 10virus particles/ml.44. The composition of claim 37 , wherein the composition comprises the replication defective adenovirus vector at a concentration of at least 10virus particles/ml.45. The composition of claim 37 , wherein the composition comprises the replication defective adenovirus vector at a concentration of at least 10virus particles/ml.46. The composition of claim 37 , wherein the composition comprises the replication defective adenovirus vector at a concentration of at least ...

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Номер записи: 78
27-02-2020 дата публикации

Transient transfection method for retroviral production

Номер: US20200063144A1
Автор: Celeste PALLANT, Conrad Vink, Eirini Vamva, Sabine Johnson
Принадлежит: GlaxoSmithKline Intellectual Property Development Ltd

The invention relates to bacterial artificial chromosomes (BAC) comprising retroviral nucleic acid sequences encoding: gag and pol proteins, and an env protein or a functional substitute thereof, wherein each of the retroviral nucleic acid sequences are arranged as individual expression constructs within the BAC. The invention also relates to uses and methods of transient transfection using said BAC.

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Номер записи: 79
27-02-2020 дата публикации

Algal based edible vaccines

Номер: US20200063152A1
Автор: Ofra Chen
Принадлежит: TransAlgae Ltd

The present invention provides edible vaccines comprising transgenic microalgae expressing at least one exogenous antigen or an intervening organism comprising the transgenic microalgae. The antigen expressing microalgae are used for oral delivery of the antigen to a target organism in its intact and functional form. The exogenous antigen, expressed in the microalgae, is characterized by exerting at least one immunogenic response in the subject consuming the vaccine.

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Номер записи: 80
11-03-2021 дата публикации

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

Номер: US20210069347A1
Автор: Farkas Michael H., Pierce Eric A., Sousa Maria E.
Принадлежит:

The present invention relates to methods and compositions for gene therapy of retinitis pigmentosa related to mutations in pre-mRNA processing factor 31 (PRPF31). 116-. (canceled)17. A method of reducing vision loss in a human subject having retinitis pigmentosa caused by mutations in PRPF31 , 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.18. The method of wherein the promoter is a CAG claim 17 , CASI claim 17 , RPE65 or VMD2 promotor.19. The method of claim 17 , wherein the vector is delivered via sub-retinal injection.20. A method of increasing expression of PRPF31 in the eye of a human subject claim 17 , 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 17 , operably linked to a promoter that drives expression in retinal pigment epithelial (RPE) cells.21. The method of wherein the promoter is a CAG claim 20 , CASI claim 20 , RPE65 or VMD2 promotor.22. The method of claim 20 , wherein the PRPF31 sequence is codon optimized.23. The method of claim 20 , wherein the vector is delivered via sub-retinal injection.24. An Adeno associated virus type 2 (AAV2) vector comprising a sequence encoding human PRPF31 claim 20 , operably linked to a promoter that drives expression in retinal pigment epithelial (RPE) cells.25. The vector of claim 24 , wherein the promotor is a CAG claim 24 , CASI claim 24 , RPE65 or VMD2 promotor.26. The vector of claim 24 , wherein the PRPF31 sequence is codon optimized.27. The vector of claim 24 , wherein the vector comprises nucleotides 1319-2818 of SEQ ID NO:34.28. A pharmaceutical preparation comprising a gene delivery system claim 24 , wherein the gene delivery ...

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Номер записи: 81
15-03-2018 дата публикации

TAT-INDUCED CRISPR/ENDONUCLEASE-BASED GENE EDITING

Номер: US20180073019A1
Автор: Khalili Kamel
Принадлежит:

Compositions and methods are provided for Tat-inducible expression of a CRISPR-associated endonuclease by a truncated HIV LTR promoter containing at least a core region and a TAR region of a HIV LTR promoter. The compositions may be used as a therapeutic treatment for the treatment and/or prevention of HIV. 1. A method of inactivating a human immunodeficiency virus (HIV) in vivo or in vitro , the method comprising administering to a subject or contacting a mammalian cell with a composition comprising an isolated nucleic acid sequence encoding a clustered regularly interspaced short palindromic repeats (CRISPR)-associated endonuclease operably linked to a truncated HIV long terminal repeat (LTR) promoter containing at least a core region and a trans activation response element (TAR) of a HIV LTR promoter.2. The method of claim 1 , wherein the isolated nucleic acid sequence further encodes at least one guide RNA that is complementary to a target nucleic acid sequence in HIV.3. The method of claim 2 , wherein the target nucleic acid sequence in HIV comprises a sequence within a coding region or non-coding region of HIV wherein the non-coding region comprises a long terminal repeat sequence of HIV.4. (canceled)5. The method of claim 3 , wherein the target nucleic acid sequence comprises a sequence within the long terminal repeat of HIV claim 3 , said sequence within the long terminal repeat of HIV comprises a sequence within the U3 claim 3 , R claim 3 , or U5 regions that excludes any sequence of the truncated HIV LTR promoter.6. (canceled)7. The method of claim 1 , wherein the mammalian cell is a latently infected cell.8. The method of claim 7 , wherein the latently infected cell is selected from the group consisting of: a CD4 T cell claim 7 , a macrophage claim 7 , a monocyte claim 7 , a gut-associated lymphoid cell claim 7 , a microglial cell claim 7 , and an astrocyte.9. (canceled)10. The method of claim 1 , wherein the mammalian cell comprises a cultured cell from ...

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Номер записи: 82
24-03-2022 дата публикации

TRANSPOSON-BASED TRANSFECTION SYSTEM FOR PRIMARY CELLS

Номер: US20220089671A1
Автор: BUNSE Mario, CLAUSS Julian, Izsvák Zsuzsanna, UCKERT Wolfgang
Принадлежит:

The present invention relates to the field of genetic engineering, in particular, to a transposon-based transfection kit suitable for transfection of primary cells, such as T cells, comprising mRNA encoding a transposase, or reagents for generating mRNA encoding said transposase, as well as minicircle DNA comprising the transposon. The invention also relates to a nucleic acid, preferably, a DNA minicircle, comprising a transposon, wherein the transposon encodes a protein and at least one miRNA, wherein the sequences encoding the miRNA are located in an intron and expression of the protein and the miRNA is regulated by the same promoter. The invention also provides a population of cells obtainable with the method of the invention. Methods of transfection are also provided, as well as medical use, e.g. in immunotherapy, in particular, in adoptive T cell therapy or T cell receptor (TCR) or chimeric antigen receptor (CAR) gene therapy. 1. A kit comprising{'claim-text': ['(i) mRNA encoding said transposase; or', '(ii) DNA encoding said transposase functionally linked to a promoter, wherein the kit optionally further comprises reagents suitable for in vitro transcription, comprising ribonucleotide triphosphates, a buffer suitable for transcription, and a RNA polymerase suitable for transcription; and'], '#text': 'a) a nucleic acid encoding a transposase capable of mobilizing a transposon, wherein the nucleic acid is selected from the group comprising'}b) minicircle DNA comprising said transposon, wherein the transposon encodes at least one protein and/or at least one miRNA, wherein expression of the protein and/or the miRNA is regulated by a promoter.2. The kit of claim 1 , wherein the nucleic acid encoding the transposase is mRNA.3. The kit of any of the preceding claims claim 1 , wherein the transposase is selected from the group of class II transposable elements comprising piggyBac claim 1 , Tol2 and MI/mariner-type transposons comprising Frog Prince and Sleeping ...

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Номер записи: 83
07-03-2019 дата публикации

METHODS FOR MAKING FULLY HUMAN BISPECIFIC ANTIBODIES USING A COMMON LIGHT CHAIN

Номер: US20190071519A1
Автор: DAVIS Samuel, MACDONALD Lynn, MCWHIRTER John, MURPHY Andrew J., STEVENS Sean
Принадлежит:

A genetically modified mouse is provided, wherein the mouse expresses an immunoglobulin light chain repertoire characterized by a limited number of light chain variable domains. Mice are provided that express just one or a few immunoglobulin light chain variable domains from a limited repertoire in their germline. Methods for making bispecific antibodies having universal light chains using mice as described herein, including human light chain variable regions, are provided. Methods for making human variable regions suitable for use in multispecific binding proteins, e.g., bispecific antibodies, and host cells are provided. Bispecific antibodies capable of binding first and second antigens are provided, wherein the first and second antigens are separate epitopes of a single protein or separate epitopes on two different proteins are provided. 120-. (canceled)21. A method of making a bispecific antigen-binding protein comprising: [ wherein the first mouse is homozygous or heterozygous in its germline genome for a single rearranged human immunoglobulin kappa light chain variable region sequence comprising a human Vκ gene segment and a human Jκ gene segment, wherein all immunoglobulin kappa light chains expressed by B cells of the first mouse comprise immunoglobulin kappa light chain variable domains expressed from the single rearranged human immunoglobulin kappa light chain variable region sequence or a somatically hypermutated version thereof, and wherein the first mouse lacks endogenous immunoglobulin Vκ and/or Jκ gene segments that are capable of rearranging to form an endogenous immunoglobulin light chain variable region sequence, and', 'wherein the first mouse expresses a first plurality of human immunoglobulin heavy chain variable domains; and, 'the first B cell is generated by a first mouse following exposure to a first epitope of a first antigen of interest,'}, wherein the second mouse is homozygous or heterozygous in its germline genome for the single ...

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Номер записи: 84
05-03-2020 дата публикации

GENE THERAPY FOR CILIOPATHIES

Номер: US20200069753A1
Автор: Beales Phillip, Hernandez Victor
Принадлежит:

There is described a vector for treating a ciliopathy such as Bardet-Biedl syndrome, wherein the vector comprises a promoter operably linked to a ciliopathy gene, wherein the vector can provide transduction of the ciliopathy gene into multiple organs, wherein the promoter is a ubiquitous promoter which can provide expression of the ciliopathy gene in the transduced organs, and wherein the ciliopathy gene encodes a functional protein corresponding to the protein that is mutated in the ciliopathy. Also described is the use of the above vector in a method of treating a ciliopathy, the method comprising administering a therapeutically effective amount of the vector to a patient suffering from a ciliopathy. 1. A vector for treating a ciliopathy , wherein the vector comprises a promoter operably linked to a ciliopathy gene , wherein the vector can provide transduction of the ciliopathy gene into multiple organs , wherein the promoter is a ubiquitous promoter which can provide expression of the ciliopathy gene in the transduced organs , and wherein the ciliopathy gene encodes a functional human protein corresponding to the protein that is mutated in the ciliopathy.2. A vector according to claim 1 , wherein the vector is an adeno-associated viral (AAV) vector or a lentiviral vector.3. A vector according to claim 1 , wherein the vector is an AAV vector.4. A vector according to claim 1 , wherein the vector is selected from AAV8 claim 1 , AAV9 claim 1 , AAV vectors pseudotyped with the capsid proteins from AAV8 or AAV9 claim 1 , AAV-PHP.A claim 1 , AAV-PHP.B claim 1 , AAV9.47 claim 1 , AAV-B1 claim 1 , AAV8(Y733F) or AAV2-TT.5. A vector according to claim 1 , wherein the vector is an AAV8 vector claim 1 , an AAV9 vector claim 1 , or an AAV vector which has been pseudotyped with the capsid proteins from AAV8 or AAV9.6. A vector according to claim 1 , wherein the promoter is selected from the short elongation factor promoter (EFS) claim 1 , CAG promoter claim 1 , cytomegalovirus ...

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Номер записи: 85
07-03-2019 дата публикации

COMPOSITIONS USEFUL IN TREATMENT OF ORNITHINE TRANSCARBAMYLASE (OTC) DEFICIENCY

Номер: US20190071651A1
Автор: Wang Lili, Wilson James M.
Принадлежит:

Viral vectors comprising engineered hOTC DNA and RNA sequences are provided which when delivered to a subject in need thereof are useful for treating hyperammonemia, ornithine transcarbamylase transcarbamylase deficiency and symptoms associated therewith. Also provided are methods of using hOTC for treatment of liver fibrosis and/or cirrhosis in OTCD patients by administering hOTC. 1. A recombinant viral vector comprising a nucleic acid sequence encoding human ornithine transcarbamylase (hOTC) and expression control sequences which direct expression of hOTC in a liver cell , wherein the hOTC nucleic acid sequence is less than 80% identical to the wild-type hOTC sequence over the mature sequence or full length hOTC of SEQ ID NO: 1 , and expresses a functional hOTC , wherein said hOTC nucleic acid sequence is SEQ ID NO: 9 or a nucleic acid sequence at least 96 to 99% identical thereto.2. A recombinant viral vector comprising a nucleic acid sequence encoding hOTC and expression control sequences which direct expression of hOTC in a liver cell , wherein the hOTC nucleic acid sequence expresses a functional hOTC , wherein said hOTC nucleic acid sequence is SEQ ID NO: 3. This is a continuation of U.S. patent application Ser. No. 15/122,853, filed Aug. 31, 2016, which is a national stage application under 35 U.S.C. 371 of PCT/US2015/019513, filed on Mar. 9, 2015, now expired, which claims the benefit of U.S. Patent Application No. 61/950,157, filed Mar. 9, 2014, now expired. These applications are incorporated by reference in their entirety.This invention was made with support under grant Nos. P01-HD057247, P01-HL059407, and P30-DK047757 awarded by the National Institutes of Health. The US government has certain rights in the invention.Ornithine transcarbamylase (OTC) deficiency accounts for nearly half of all cases of inborn errors of urea synthesis, with a prevalence estimated to be at least 1 in 15,000. Urea cycle defects put patients at risk of life threatening ...

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Номер записи: 86
07-03-2019 дата публикации

Engineered CRISPR-Cas9 Nucleases

Номер: US20190071657A1
Автор: Joung J. Keith, Kleinstiver Benjamin, Pattanayak Vikram
Принадлежит:

Engineered CRISPR-Cas9 nucleases with improved specificity and their use in genomic engineering, epigenomic engineering, genome targeting, and genome editing. 1Streptococcus pyogenes. An isolated Cas9 (SpCas9) protein , with mutations at one or both of Q695 and Q926 , and optionally one or more of a nuclear localization sequence , cell penetrating peptide sequence , and/or affinity tag.2. The isolated protein of claim 1 , comprising all four of the following mutations:N497A, R661A, Q695A, and Q926A.3. The isolated protein of claim 1 , comprising mutations at one or both of Q695 and Q926 claim 1 , and optionally one claim 1 , two claim 1 , three claim 1 , four claim 1 , or all five of L169 claim 1 , Y450 claim 1 , N497 claim 1 , R661 claim 1 , and D1135.4. The isolated protein of further comprising mutations at one claim 2 , two claim 2 , three claim 2 , four claim 2 , or all five of L169 claim 2 , Y450 claim 2 , N497 claim 2 , R661 claim 2 , and D1135.5. The isolated protein of claim 1 , further comprising one or more of the following mutations: D1135E; D1135V; D1135V/R1335Q/T1337R (VQR variant); D1135E/R1335Q/T1337R (EQR variant); D1135V/G1218R/R1335Q/T1337R (VRQR variant); or D1135V/G1218R/R1335E/T1337R (VRER variant).6. The isolated protein of claim 1 , further comprising one or more mutations that decrease nuclease activity selected from the group consisting of mutations at D10 claim 1 , E762 claim 1 , D839 claim 1 , H983 claim 1 , or D986; and at H840 or N863.7. The isolated protein of claim 6 , wherein the mutations that decrease nuclease activity are:(i) D10A or D10N, and(ii) H840A, H840N, or H840Y.8. A fusion protein comprising the isolated protein of claim 1 , fused to a heterologous functional domain claim 1 , with an optional intervening linker claim 1 , wherein the linker does not interfere with activity of the fusion protein.9. The fusion protein of claim 8 , wherein the heterologous functional domain is a transcriptional activation domain.10. The ...

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Номер записи: 87
05-03-2020 дата публикации

Mva vaccine for delivery of a ul128 complex and preventing cmv infection

Номер: US20200069791A1
Автор: Don J. Diamond, Felix WUSSOW
Принадлежит: CITY OF HOPE

In one embodiment, an expression system for expressing a UL128 complex is provided herein. The expression system may include a bacterial artificial chromosome (BAC) construct, wherein the BAC construct comprises a viral vector inserted with a set of DNA sequences that encode a UL128 complex. In another embodiment, a vaccine composition for preventing HCMV infection is provided. The vaccine composition may include a viral or bacterial vector capable of expressing a UL128 complex and a pharmaceutically acceptable carrier, adjuvant, additive or combination thereof or additional vector expressing a protein adjuvant. The viral vector may be an MVA and the UL128 complex includes five HCMV proteins or antigenic fragments thereof: UL128, UL130, UL131A, gL, and gH. In some embodiments, the viral vector is further inserted with one or more additional DNA sequences that encode one or more additional HCMVHCMV proteins or antigenic fragments thereof such as pp65, gB or both, or such as gM/gN or gO.

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Номер записи: 88
07-03-2019 дата публикации

Lineage reporter synthetic chromosomes and methods of use

Номер: US20190071738A1
Автор: Amy Greene, Edward Perkins
Принадлежит: Synploid Biotek LLC

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.

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Номер записи: 89
24-03-2022 дата публикации

PROCESS FOR THE MANIPULATION OF NUCLEIC ACIDS

Номер: US20220090143A1
Автор: Faridmoayer Amirreza, KOWARIK Michael Thomas, LIPOWSKY Gerd Martin, SERVENTI Fabio
Принадлежит: GLAXOSMITHKLINE BIOLOGICALS SA

The present invention discloses a process for engineering a host cell comprising the steps of; a) integrating a first polynucleotide cassette including a first selection marker flanked by a first pair of recombination sites; b) removing the first selection marker by the action of a recombinase which recognises the first pair of recombination sites; c) integrating a second polynucleotide cassette including a second selection marker flanked by a second pair of recombination sites; and d) removing the second selection marker by the action of a recombinase which recognises the second pair of recombination sites. Also disclosed is a host cell genome polynucleotide comprising a first recombinantly engineered region and a second recombinantly engineered region, wherein a first single recombination site is adjacent to the first recombinantly engineered region, and a second single recombination site is adjacent to the second recombinantly engineered region. 125.-. (canceled)26. A method of removing at least two portions of insert nucleic acid from a genomic polynucleotide in a host cell , said method comprising the steps of:a) preparing the genomic polynucleotide comprising a first insert nucleic acid which is flanked by a pair of first recombination sites in the same orientation which are identical to each other and have a first nucleic acid sequence;b) exposing the genomic polynucleotide of step a) to a recombinase that recognises the first recombination sites such that the identical recombination sites recombine resulting in the excision of the first insert nucleic acid and one of the first recombination sites;c) inserting into the genomic polynucleotide of step b) a second insert nucleic acid flanked by a pair of second recombination sites in the same orientation wherein the second recombination sites are identical to each other and have a second nucleic acid sequence which shares 70-98% sequence identity with the first nucleic acid sequence; andd) exposing the genomic ...

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Номер записи: 90
05-03-2020 дата публикации

PARTICLE FOR THE ENCAPSIDATION OF A GENOME ENGINEERING SYSTEM

Номер: US20200071720A1
Автор: BOUILLE Pascale, GAYON Régis, ICHE Alexandra, LAMOUROUX Lucille
Принадлежит:

The present invention relates to a retroviral particle comprising a protein derived from the Gag polyprotein, an envelope protein, optionally an integrase and at least two encapsidated non-viral RNAs, the encapsidated non-viral RNAs each comprising an RNA sequence of interest bound to an encapsidation sequence, each encapsidation sequence being recognized by a binding domain introduced into the protein derived from the Gag polyprotein and/or into the integrase, and at least one of said sequences of interest of the encapsidated non-viral RNAs comprises a part coding a nuclease. 1. Retroviral particle comprising a protein derived from the Gag polyprotein , an envelope protein , optionally an integrase and at least two encapsidated non-viral RNAs , the encapsidated non-viral RNAs each comprising an RNA sequence of interest bound to an encapsidation sequence , each encapsidation sequence being recognized by a binding domain introduced into the protein derived from the Gag polyprotein and/or into the integrase , and at least one of said sequences of interest of the encapsidated non-viral RNAs comprises a part coding a nuclease.2Natronobacterium gregoryi. Particle according to claim 1 , in which the nuclease is selected from the group constituted by the nucleases associated with the CRISPR system claim 1 , the nuclease of the TALEN system claim 1 , the nuclease of the Zn Finger system and the Argonaute (NgAgo) nuclease.3. Retroviral particle according to claim 1 , in which the sequence of interest of at least two encapsidated non-viral RNAs comprises a part coding a nuclease.4. Retroviral particle according to claim 1 , in which the at least two encapsidated non-viral RNAs differ by their sequence of interest.5. Retroviral particle according to claim 4 , in which:the sequence of interest of at least one RNA comprises a part coding a nuclease and a part coding a DNA recognition system at 3′, andthe sequence of interest of at least one RNA comprises a part coding a nuclease ...

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Номер записи: 91
18-03-2021 дата публикации

NOVEL CRISPR-ASSOCIATED (CAS) PROTEIN

Номер: US20210079367A1
Автор: Carter Matthew Merrill, Donohoue Paul Daniel
Принадлежит:

A new CRISPR-associated (Cas) protein, termed “CasM,” is described, as well as polynucleotides encoding the same and methods of using CasM for site-specific genome engineering. CasM proteins are capable of targeting and cleaving single-stranded RNA. 1. A Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR)-associated (Cas) nucleoprotein complex , wherein the nucleoprotein complex comprises a Cas protein having at least 95 percent sequence identity to the amino acid sequence of SEQ ID NO: 40; anda cognate nucleic acid guide comprising a repeat sequence and a spacer sequence, wherein the repeat sequence and the spacer sequence do not naturally occur together, wherein the Cas protein and the cognate nucleic acid guide form the nucleoprotein complex, and wherein the nucleoprotein complex is capable of site-directed binding to a target nucleic acid sequence.2. The Cas nucleoprotein complex of claim 1 , wherein the cognate nucleic acid guide comprises a crRNA.3. The Cas nucleoprotein complex of claim 1 , wherein the Cas protein comprises an N- or C-terminal nuclear localization signal sequence (NLS).4. The Cas nucleoprotein complex of claim 1 , wherein the cognate nucleic acid guide comprises a modified base analog.5. The Cas nucleoprotein complex of claim 1 , wherein the sequence identity is calculated using BLAST computer program with default parameters.6. A eukaryotic cell comprising the Cas nucleoprotein complex of .7. The eukaryotic cell of claim 6 , further comprising one or more additional Cas nucleoprotein complexes.8. The eukaryotic cell of claim 7 , wherein the eukaryotic cell is a human cell.9. The Cas nucleoprotein complex of claim 1 , wherein the Cas protein is catalytically active.10. The Cas nucleoprotein complex of claim 1 , wherein the Cas protein is catalytically inactive.11. The Cas nucleoprotein complex of claim 1 , wherein the Cas protein comprises a fusion protein.12. An expression cassette comprising:a polynucleotide encoding:a ...

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Номер записи: 92
18-03-2021 дата публикации

METHODS TO PRODUCE CHIMERIC ADENO-ASSOCIATED VIRUS/BOCAVIRUS PARVOVIRUS

Номер: US20210079421A1
Автор: Engelhardt John F., Qiu Jianming, Yan Ziying
Принадлежит:

A method of preparing a chimeric virus comprising bocavirus capsid protein (VP) and a recombinant adeno-associated (AAV) viral genome, and isolated mutant bocavirus genomes, are provided. 1. A method of preparing a chimeric virus comprising bocavirus capsid protein (VP) and a recombinant adeno-associated (AAV) viral genome , comprising: i) does not express one or more of bocavirus NS1, NS2, NS3 or NS4;', 'ii) expresses bocavirus VP1, VP2 and VP3; and', 'iii) expresses bocavirus NP1;, 'a) providing a mutant bocavirus genome that when introduced to cells;'}b) introducing the mutant bocavirus genome and a rAAV vector to cells which do not express one or more of bocavirus NS1, NS2, NS3 or NS4, thereby producing bocavirus VP, bocavirus NP1 and a rAAV genome; andc) collecting chimeric rAAV/bocavirus.2. The method of wherein the rAAV vector comprises an expression cassette encoding a heterologous gene product.3. The method of wherein the gene product encodes a therapeutic protein.4. The method of wherein the rAAV genome is a rAAV-1 claim 1 , rAAV-2 claim 1 , rAAV-3 claim 1 , rAAV-4 claim 1 , rAAV-5 claim 1 , rAAV-6 claim 1 , rAAV-7 claim 1 , rAAV-8 or rAAV-9 genome.5. The method of wherein the gene product is a viral claim 2 , bacterial claim 2 , tumor claim 2 , parasite claim 2 , or fungal antigen.6. The method of wherein the gene product is cystic fibrosis transmembrane conductance regulator claim 2 , β-globin claim 2 , γ-globin claim 2 , tyrosine hydroxylase claim 2 , glucocerebrosidase claim 2 , aryl sulfatase A claim 2 , factor VIII claim 2 , dystrophin claim 2 , alpha 1-antitrypsin claim 2 , surfactant protein SP-D claim 2 , SP-A or SP-C claim 2 , C1 inhibitor gene claim 2 , C1-INH gene claim 2 , SERPING gene claim 2 , erythropoietin claim 2 , HBoV protein claim 2 , influenza virus protein claim 2 , RSV protein claim 2 , a neutralizing antibody or an antigen binding fragment thereof claim 2 , SARS virus protein claim 2 , or a cytokine claim 2 , e.g. claim 2 , IFN- ...

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Номер записи: 93
14-03-2019 дата публикации

MICE EXPRESSING A LIMITED IMMUNOGLOBULIN LIGHT CHAIN REPERTOIRE

Номер: US20190077884A1
Автор: MACDONALD Lynn, MCWHIRTER John, MURPHY Andrew J., STEVENS Sean
Принадлежит:

A genetically modified mouse is provided, wherein the mouse expresses an immunoglobulin light chain repertoire characterized by a limited number of light chain variable domains. Mice are provided that present a choice of two human light chain variable gene segments such that the immunoglobulin light chains expresses by the mouse comprise one of the two human light chain variable gene segments. Methods for making bispecific antibodies having universal light chains using mice as described herein, including human light chain variable regions, are provided. Methods for making human variable regions suitable for use in multispecific binding proteins, e.g., bispecific antibodies, and host cells are provided. 130-. (canceled)31. A method for generating a human immunoglobulin heavy or light chain variable domain sequence comprising the steps of: (i) exactly two unrearranged human immunoglobulin Vκ gene segments and five unrearranged human immunoglobulin Jκ gene segments operably linked to a mouse immunoglobulin light chain constant region sequence at the endogenous kappa light chain loci of the mouse, wherein the two unrearranged human immunoglobulin Vκ gene segments are a human Vκ1-39 gene segment and a human Vκ3-20 gene segment; and', {'sub': H', 'H', 'H, 'claim-text': 'wherein the unrearranged human immunoglobulin heavy chain and kappa light chain gene segments of the genetically modified mouse are capable of rearranging and encoding human immunoglobulin variable domains of an antibody, wherein the genetically modified mouse does not comprise endogenous immunoglobulin Vκ or Jκ gene segments that are capable of rearranging to form an immunoglobulin light chain variable region sequence; and', '(ii) one or more unrearranged human immunoglobulin Vgene segments, one or more unrearranged human immunoglobulin Dgene segments, and one or more unrearranged human immunoglobulin Jgene segments operably linked to a mouse immunoglobulin heavy chain constant region sequence at the ...

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Номер записи: 94
22-03-2018 дата публикации

VIRAL VECTORS FOR THE TREATMENT OF RETINAL DYSTROPHY

Номер: US20180080046A1
Автор: BIGELOW Chad Eric, CHOI Vivian, DRYJA Thaddeus Peter, POLICE Seshidhar Reddy
Принадлежит: NOVARTIS AG

The present invention relates to viral vectors that are capable of delivering a heterologous gene to the retina and in particular delivering RLBP1 to RPE and Müller cells of the retina. The invention also relates nucleic acids useful for producing viral vectors, compositions comprising the viral vectors and uses of the compositions and viral vectors. The invention also relates to methods of delivering and/or expressing a heterologous gene to the retina, improving the rate of dark adaption in a subject and treating RLBP1-associated retinal dystrophy. 114-. (canceled)15. A method of expressing a heterologous gene in retinal cells , the method comprising contacting said retinal cells with a viral vector comprising: a) an AAV2 or AAV8 capsid , and b) a vector genome comprising a retinaldehyde binding protein 1 (RLBP1) promoter comprising the nucleotide sequence of SEQ ID NO:3 or SEQ ID NO: 10 operably linked to the heterologous gene.16. The method of claim 15 , wherein the retinal cells are RPE cells and Müller cells.17. The method of claim 16 , wherein the vector genome is a self-complementary genome.18. The method of claim 15 , wherein the heterologous gene is RLBP1.1924-. (canceled)25. A method of treating a subject having RLBP1-associated retinal dystrophy claim 15 , the method comprising administering to the subject a therapeutically effective amount of a composition comprising a viral vector comprising:a vector genome comprising the nucleotide sequences of SEQ ID NOs:5, 6, 8, and 9; andan adeno-associated virus (AAV) serotype 2 or 8 capsid.26. A method of improving the rate of dark adaption in a subject having RLBP1-associated retinal dystrophy claim 15 , the method comprising administering to the subject a therapeutically effective amount of a composition comprising a viral vector comprising:a vector genome comprising the nucleotide sequences of SEQ ID NOs:5, 6, 8, and 9; andan adeno-associated virus (AAV) serotype 2 or 8 capsid.27. A method of expressing an ...

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Номер записи: 95
31-03-2022 дата публикации

Nucleic acid therapeutics for genetic disorders

Номер: US20220096525A1
Автор: Andrey Jury Zarur, Christian Matranga, James Robbins Abshire, Katherine J. Williams, Kelsey A. Haugh
Принадлежит: Greenlight Biosciences Inc

Provided herein, are compositions based on retroviruses (e.g., lentiviruses) comprising one or more nucleic acid molecules encoding retroviral Pol polyprotein components and a nucleic acid molecule comprising one or more transgene sequences flanked by long terminal repeat sequences, for delivery of the one or more transgenes to a target cell ex vivo or in vivo. The compositions are useful for delivering to a target cell (e.g., hematopoietic stem cells (HSCs), liver cells, ocular cells, muscle cells, epithelial cells, T cells, etc.) and/or stably expressing any transgene (e.g., beta-globin, Factor VIII, RP GTPase regulator (RPGR), dystrophin, cystic fibrosis transmembrane conductance regulator (CFTR), a chimeric antigen receptor, etc.) with a biological effect to treat and/or ameliorate the symptoms associated with any disorder related to gene expression (e.g., sickle cell disease, beta-thalassemia, haemophilia B, retinitis pigmentosa, Duchenne muscular dystrophy, cystic fibrosis, cancer, etc.).

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Номер записи: 96
31-03-2022 дата публикации

METHODS FOR MODIFYING THE GROWTH RATE OF A CELL

Номер: US20220098598A1
Автор: Cohen-Kupiec Rachel, Tuller Tamir, Zur Hadas
Принадлежит:

Genetically modified cells with at least one codon substituted to a synonymous codon, and with modified replicative fitness as compared to the unmodified cell, wherein a slower translating synonymous codon increases replicative fitness and a faster translating codon decreased replicative fitness are provided. Further, vaccine composition comprising those cells as well as methods for modifying replicative fitness of a cell are provided. 1. A method of modifying replicative fitness in a cell , the method comprising modifying ribosome density upstream of a ribosome backup on at least one translating sequence in said cell , wherein increasing ribosome density upstream of a ribosome backup decreases replicative fitness and decreasing ribosome density upstream of a ribosome backup increased replicative fitness , thereby modifying replicative fitness in a cell.2. The method of claim 1 , wherein said ribosome backup comprises a region upstream of said backup with a ribosome density at least 5% higher than a ribosome density downstream of said backup.3. The method of claim 1 , wherein said ribosome backup comprises a slowly translating codon.4. The method of claim 3 , wherein said slowly translating codon translates at a slower rate than the average translational rate of codons 11 to 50 of said translating sequence.5. The method of claim 3 , wherein said slowly translating codon is downstream of codon 50 from a translational start site of said translating sequence.6. The method of claim 1 , wherein said modifying ribosome density does not substantially decrease the translation efficiency of said translating sequence.7. The method of claim 6 , wherein a substantial decrease is a decrease of at least 5%.8. The method of claim 6 , wherein said modifying ribosome density comprises substituting a codon with a different codon and further comprises determining whether said substituting would reduce translation efficiency below said threshold claim 6 , and wherein said determining ...

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Номер записи: 97
12-03-2020 дата публикации

NOVEL CRISPR-ASSOCIATED (CAS) PROTEIN

Номер: US20200080068A1
Автор: Carter Matthew Merrill, Donohoue Paul Daniel
Принадлежит:

A new CRISPR-associated (Cas) protein, termed “CasM,” is described, as well as polynucleotides encoding the same and methods of using CasM for site-specific genome engineering. CasM proteins are capable of targeting and cleaving single-stranded RNA. 132-. (canceled)33. A method of directing a Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR)-associated (Cas) protein to a selected nucleic acid target sequence , the method comprising: a Cas protein, wherein the Cas protein is selected from the group consisting of SEQ ID NO:37, SEQ ID NO:39, and SEQ ID NO:44, an amino acid sequence having at least 90 percent sequence identity to SEQ ID NO:37, an amino acid sequence having at least 90 percent sequence identity to SEQ ID NO:39, and an amino acid sequence having at least 90 percent sequence identity to SEQ ID NO:44, and', 'a cognate nucleic acid-targeting nucleic acid (NATNA)., 'contacting the selected nucleic acid target sequence with a nucleoprotein complex comprising'}34. The method of claim 33 , wherein the nucleic acid target sequence comprises DNA.35. (canceled)36. (canceled)37. The method of claim 33 , wherein the method is performed in a cell.38. The method of claim 37 , wherein the cell is a eukaryotic cell.39. The method of claim 37 , wherein the cell constitutively expresses the Cas protein.40. (canceled)41. The method of claim 33 , wherein the Cas protein is fused to a protein.42. The method of claim 41 , wherein the protein comprises an enzyme.43. The method of claim 33 , wherein the NATNA comprises RNA.44. The method of claim 43 , wherein the NATNA comprises DNA.45. The method of claim 33 , wherein the Cas protein comprises SEQ ID NO:39 or an amino acid sequence having at least 90 percent sequence identity to SEQ ID NO:39.46. The method of claim 33 , wherein the nucleoprotein complex is capable of site-directed binding to the nucleic acid target sequence.47. The method of claim 46 , wherein the Cas protein is catalytically active claim 46 , ...

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Номер записи: 98
12-03-2020 дата публикации

CIRCULAR RNA FOR TRANSLATION IN EUKARYOTIC CELLS

Номер: US20200080106A1
Автор: ANDERSON Daniel G., Kowalski Piotr S., Wesselhoeft Robert Alexander
Принадлежит:

Disclosed are methods and constructs for engineering circular RNA. Disclosed is a vector for making circular RNA, said vector comprising the following elements operably connected to each other and arranged in the following sequence: 1. A vector for making circular RNA , said vector comprising the following elements operably connected to each other and arranged in the following sequence:a) a 5′ homology arm,b) a 3′ Group I intron fragment containing a 3′ splice site dinucleotide,c) a 5′ spacer sequence,d) an internal ribosome entry site (IRES),e) a protein coding region or noncoding region,f) a 3′ spacer sequence,g) a 5′ Group I intron fragment containing a 5′ splice site dinucleotide, andh) a 3′ homology arm,said vector allowing production of a circular RNA that is translatable or biologically active inside eukaryotic cells.2. (canceled)3Cyanobacterium Anabaena. The vector of claim 1 , wherein the 3′ Group I intron fragment and the 5′ Group I intron fragment are from a sp. pre-tRNA-Leu gene.4. The vector of claim 1 , wherein the 3′ Group I intron fragment and the 5′ Group I intron fragment are from a T4 phage Td gene.5S. cerevisiaeS. cerevisiae. The vector of claim 1 , wherein the IRES is selected from an IRES sequence of Taura syndrome virus claim 1 , Triatoma virus claim 1 , Theiler's encephalomyelitis virus claim 1 , simian Virus 40 claim 1 , Solenopsis invicta virus 1 claim 1 , Rhopalosiphum padi virus claim 1 , Reticuloendotheliosis virus claim 1 , fuman poliovirus 1 claim 1 , Plautia stali intestine virus claim 1 , Kashmir bee virus claim 1 , Human rhinovirus 2 claim 1 , Homalodisca coagulata virus-1 claim 1 , Human Immunodeficiency Virus type 1 claim 1 , Homalodisca coagulata virus-1 claim 1 , Himetobi P virus claim 1 , Hepatitis C virus claim 1 , Hepatitis A virus claim 1 , Hepatitis GB virus claim 1 , foot and mouth disease virus claim 1 , Human enterovirus 71 claim 1 , Equine rhinitis virus claim 1 , Ectropis obliqua picorna-like virus claim 1 , ...

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Номер записи: 99
12-03-2020 дата публикации

NOVEL CRISPR ENZYMES AND SYSTEMS

Номер: US20200080112A1
Автор: CHOUDHURY Sourav, HEIDENREICH Matthias, SCOTT David Arthur, YAN Winston Xia, Zhang Feng
Принадлежит:

Embodiments disclosed herein are directed to engineered CRISPR-Cas effector proteins that comprise at least one modification compared to an unmodified CRISPR-Cas effector protein that enhances binding of the of the CRISPR complex to the binding site and/or alters editing preference as compared to wild type. In certain example embodiments, the CRISPR-Cas effector protein is a Type V effector protein. In certain other example embodiments, the Type V effector protein is Cpf1. Embodiments disclosed herein are directed to viral vectors for delivery of CRISPR-Cas effector proteins, including Cpf1. In certain example embodiments, the vectors are designed so as to allow packaging of the CRISPR-Cas effector protein within a single vector. There is also an increased interest in the design of compact promoters for packing and thus expressing larger transgenes for targeted delivery and tissue-specificity. Thus, in another aspect certain embodiments disclosed herein are directed to delivery vectors, constructs, and methods of delivering larger genes for systemic delivery. 134-. (canceled)35. A method for developing or designing a CRISPR-Cas system based therapy or therapeutic , comprising:selecting a set of target sequences for one or more loci in a target population, wherein the target sequences do not contain variants occurring above a threshold allele frequency in the target population;removing any target sequences having high frequency off-target candidates (relative to other platinum targets in the set) to define a final target sequence set;preparing a set of CRISPR-Cas systems based on the final target sequence set, wherein a number of CRISPR-Cas systems prepared is based at least in part a size of a target population.36. The method of claim 35 , further comprising;obtaining genome sequencing data of a subject to be treated; andtreating the subject with a CRISPR-Cas system selected from the set of CRISPR-Cas systems, wherein the CRISPR-Cas system selected is based at least ...

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Номер записи: 100