EA037255B1 - Новые сайты интеграции cho и их применения - Google Patents
Новые сайты интеграции cho и их применения Download PDFInfo
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- EA037255B1 EA037255B1 EA201790698A EA201790698A EA037255B1 EA 037255 B1 EA037255 B1 EA 037255B1 EA 201790698 A EA201790698 A EA 201790698A EA 201790698 A EA201790698 A EA 201790698A EA 037255 B1 EA037255 B1 EA 037255B1
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Classifications
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
- C12N15/09—Recombinant DNA-technology
- C12N15/87—Introduction of foreign genetic material using processes not otherwise provided for, e.g. co-transformation
- C12N15/90—Stable introduction of foreign DNA into chromosome
- C12N15/902—Stable introduction of foreign DNA into chromosome using homologous recombination
- C12N15/907—Stable introduction of foreign DNA into chromosome using homologous recombination in mammalian cells
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
- C12N15/09—Recombinant DNA-technology
- C12N15/63—Introduction of foreign genetic material using vectors; Vectors; Use of hosts therefor; Regulation of expression
- C12N15/79—Vectors or expression systems specially adapted for eukaryotic hosts
- C12N15/85—Vectors or expression systems specially adapted for eukaryotic hosts for animal cells
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- Health & Medical Sciences (AREA)
- Genetics & Genomics (AREA)
- Life Sciences & Earth Sciences (AREA)
- Engineering & Computer Science (AREA)
- Wood Science & Technology (AREA)
- Biomedical Technology (AREA)
- Organic Chemistry (AREA)
- Biotechnology (AREA)
- General Engineering & Computer Science (AREA)
- Chemical & Material Sciences (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Zoology (AREA)
- Molecular Biology (AREA)
- Biophysics (AREA)
- Microbiology (AREA)
- Plant Pathology (AREA)
- Biochemistry (AREA)
- General Health & Medical Sciences (AREA)
- Physics & Mathematics (AREA)
- Cell Biology (AREA)
- Mycology (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
- Preparation Of Compounds By Using Micro-Organisms (AREA)
- Peptides Or Proteins (AREA)
- Saccharide Compounds (AREA)
- Polysaccharides And Polysaccharide Derivatives (AREA)
Applications Claiming Priority (2)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| US201462067774P | 2014-10-23 | 2014-10-23 | |
| PCT/US2015/056653 WO2016064999A1 (en) | 2014-10-23 | 2015-10-21 | Novel cho integration sites and uses thereof |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| EA201790698A1 EA201790698A1 (ru) | 2018-01-31 |
| EA037255B1 true EA037255B1 (ru) | 2021-02-26 |
Family
ID=54602002
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| EA201790698A EA037255B1 (ru) | 2014-10-23 | 2015-10-21 | Новые сайты интеграции cho и их применения |
Country Status (16)
| Country | Link |
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| US (5) | US9816110B2 (enExample) |
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Families Citing this family (38)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| KR102243243B1 (ko) | 2014-10-23 | 2021-04-22 | 리제너론 파마슈티칼스 인코포레이티드 | 신규한 cho 통합 부위 및 이의 용도 |
| US10329594B1 (en) | 2015-09-03 | 2019-06-25 | CHO Plus, Inc. | Cell lines for high level production of protein-based pharmaceuticals |
| US11649449B2 (en) | 2015-09-03 | 2023-05-16 | CHO Plus, Inc. | Hybrid cell lines for high level production of a target protein |
| EP3445780A1 (en) * | 2016-04-20 | 2019-02-27 | Regeneron Pharmaceuticals, Inc. | Compositions and methods for making antibodies based on use of an expression-enhancing locus |
| TWI827531B (zh) * | 2016-04-20 | 2024-01-01 | 美商再生元醫藥公司 | 基於使用增強表現基因座之製備抗體的組成物及方法 |
| WO2018039499A1 (en) | 2016-08-24 | 2018-03-01 | Regeneron Pharmaceuticals, Inc. | Host cell protein modification |
| US11499139B2 (en) * | 2016-12-20 | 2022-11-15 | Development Center For Biotechnology | Targeted integration sites in Chinese hamster ovary cell genome |
| WO2018148196A1 (en) * | 2017-02-07 | 2018-08-16 | Sigma-Aldrich Co. Llc | Stable targeted integration |
| CA3053304A1 (en) * | 2017-02-17 | 2018-08-23 | Lonza Ltd. | Mammalian cells for producing adeno-associated viruses |
| WO2018175153A1 (en) * | 2017-03-19 | 2018-09-27 | Applied Stemcell, Inc. | Novel integration sites and uses thereof |
| EP3665291B1 (en) * | 2017-08-11 | 2021-10-06 | Boehringer Ingelheim International GmbH | Integration sites in cho cells |
| CN107723276B (zh) * | 2017-11-02 | 2021-08-13 | 上海交通大学 | 一种稳定高表达目标产物的细胞株的构建方法和试剂盒 |
| IL275462B1 (en) * | 2017-12-22 | 2025-09-01 | Genentech Inc | Targeted integration of nucleic acids |
| EP3728581B1 (en) * | 2017-12-22 | 2024-10-09 | Bio-Rad Laboratories, Inc. | Controlling phenotype of organisms with crispr/cas gene targeting |
| CN109207432B (zh) * | 2018-10-30 | 2021-08-03 | 江南大学 | 一种cho细胞基因组内nw_006883358-1稳定表达蛋白质的应用 |
| CN109295092B (zh) * | 2018-10-30 | 2021-06-29 | 江南大学 | 一种cho细胞基因组内nw_003613638-1稳定表达蛋白质的应用 |
| CN109136193B (zh) * | 2018-10-30 | 2021-07-06 | 江南大学 | 一种cho细胞基因组内nw_006884764-1稳定表达蛋白质的应用 |
| CN109337927B (zh) * | 2018-10-30 | 2021-07-06 | 江南大学 | 一种cho细胞基因组内nw_006880285-1稳定表达蛋白质的应用 |
| CN109321604B (zh) * | 2018-10-30 | 2021-07-06 | 江南大学 | 一种cho细胞基因组内nw_006882077-1稳定表达蛋白质的应用 |
| CN109295093B (zh) * | 2018-10-30 | 2021-08-03 | 江南大学 | 一种cho细胞基因组内nw_006882456-1稳定表达蛋白质的应用 |
| AU2019403279A1 (en) * | 2018-12-21 | 2021-07-01 | Genentech, Inc. | Targeted integration of nucleic acids |
| EP3950938A4 (en) | 2019-04-02 | 2023-01-18 | Chugai Seiyaku Kabushiki Kaisha | METHOD FOR INTRODUCING A FOREIGN TARGET-SPECIFIC GENE |
| AU2020308002A1 (en) | 2019-06-26 | 2022-01-20 | Genentech, Inc. | Randomized configuration targeted integration of nucleic acids |
| EP3901266A1 (en) * | 2020-04-22 | 2021-10-27 | LEK Pharmaceuticals d.d. | Super-enhancers for recombinant gene expression in cho cells |
| EP4158041A4 (en) * | 2020-06-02 | 2024-09-25 | Catalent Pharma Solutions, LLC | CELL LINES WITH MULTIPLE DOCKING ZONES FOR GENE INSERTION |
| JP2023532665A (ja) * | 2020-06-24 | 2023-07-31 | ジェネンテック， インコーポレイテッド | 核酸の標的化組み込み |
| CA3197730A1 (en) | 2020-10-15 | 2022-04-21 | F. Hoffman-La Roche Ag | Nucleic acid constructs for va rna transcription |
| CN113755447B (zh) * | 2021-09-23 | 2022-04-29 | 云舟生物科技(广州)股份有限公司 | 一种用于生产腺病毒的293a细胞株及其制备与应用 |
| EP4419698A1 (en) | 2021-10-18 | 2024-08-28 | Regeneron Pharmaceuticals, Inc. | Mammalian cells comprising integrated cas9 genes to produce stable integration sites, and mammalian cells comprising stable integration sites and other sites |
| AU2022373653A1 (en) | 2021-10-18 | 2024-05-02 | Regeneron Pharmaceuticals, Inc. | Eukaryotic cells comprising adenovirus-associated virus polynucleotides |
| US20230304062A1 (en) | 2021-10-18 | 2023-09-28 | Regeneron Pharmaceuticals, Inc. | Controlled transcription of polynucleotides |
| AU2023213996A1 (en) * | 2022-01-28 | 2024-06-27 | The Trustees Of Columbia University In The City Of New York | Genetic modifications for xenotransplantation |
| CN117305334B (zh) * | 2022-06-21 | 2024-11-08 | 深圳太力生物技术有限责任公司 | 靶向整合细胞及其制备方法、生产目标基因表达产物的方法 |
| US20240167984A1 (en) | 2022-11-18 | 2024-05-23 | Regeneron Pharmaceuticals, Inc. | Methods for detecting and determining protein structures and stability in fluids, including biological fluids |
| CN118726475B (zh) * | 2023-03-28 | 2025-07-08 | 深圳太力生物技术有限责任公司 | 核酸、重组载体、靶向整合细胞、基因表达方法和应用 |
| WO2024211287A1 (en) * | 2023-04-03 | 2024-10-10 | Seagen Inc. | Production cell lines with targeted integration sites |
| US20250122482A1 (en) | 2023-09-07 | 2025-04-17 | Regeneron Pharmaceuticals, Inc. | Production and purification of covalently surface modified adeno-associated virus |
| WO2025111473A1 (en) | 2023-11-21 | 2025-05-30 | Regeneron Pharmaceuticals, Inc. | Covalently surface modified adeno-associated virus production by in vitro conjugation, and purification of covalently surface modified adeno-associated virus |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2008151219A1 (en) * | 2007-06-04 | 2008-12-11 | Regeneron Pharmaceuticals, Inc. | Enhanced expression and stability regions |
| WO2012138887A1 (en) * | 2011-04-05 | 2012-10-11 | The Scripps Research Institute | Chromosomal landing pads and related uses |
Family Cites Families (13)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US4634665A (en) | 1980-02-25 | 1987-01-06 | The Trustees Of Columbia University In The City Of New York | Processes for inserting DNA into eucaryotic cells and for producing proteinaceous materials |
| US4656134A (en) | 1982-01-11 | 1987-04-07 | Board Of Trustees Of Leland Stanford Jr. University | Gene amplification in eukaryotic cells |
| US6689606B2 (en) | 1998-07-21 | 2004-02-10 | M.L. Laboratories Plc | Polynucleotide |
| US6521419B1 (en) * | 1998-09-22 | 2003-02-18 | Kanakaraju Koduri | Expression vectors containing hot spot for increased recombinant protein expression in transfected cells |
| AU2002254529A1 (en) | 2001-04-04 | 2002-10-21 | The Government Of The United States Of America, As Represented By The Secretary Of The Department Of | Nucleic acids for transgene expression |
| US20040045043A1 (en) * | 2002-05-20 | 2004-03-04 | Finney Robert E. | Compositions and methods for generating conditional knockouts |
| US8673589B2 (en) | 2002-05-29 | 2014-03-18 | Regeneron Pharmaceuticals, Inc. | Inducible eukaryotic expression system |
| CN102625655B (zh) * | 2008-12-04 | 2016-07-06 | 桑格摩生物科学股份有限公司 | 使用锌指核酸酶在大鼠中进行基因组编辑 |
| PL2699264T3 (pl) | 2011-04-20 | 2018-08-31 | Medimmune, Llc | Przeciwciała i inne cząsteczki wiążące B7-H1 i PD-1 |
| DK3489366T3 (da) * | 2011-06-01 | 2020-02-24 | Prec Biosciences Inc | Fremgangsmåder og produkter til produktion af manipulerede mammale cellelinier med forstærkede transgener |
| CA2852955C (en) * | 2011-10-27 | 2021-02-16 | Sangamo Biosciences, Inc. | Methods and compositions for modification of the hprt locus |
| BR112014027813A2 (pt) | 2012-05-07 | 2017-08-08 | Dow Agrosciences Llc | métodos e composições para integração de transgenes direcionada mediada por nuclease |
| KR102243243B1 (ko) | 2014-10-23 | 2021-04-22 | 리제너론 파마슈티칼스 인코포레이티드 | 신규한 cho 통합 부위 및 이의 용도 |
-
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-
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-
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Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2008151219A1 (en) * | 2007-06-04 | 2008-12-11 | Regeneron Pharmaceuticals, Inc. | Enhanced expression and stability regions |
| WO2012138887A1 (en) * | 2011-04-05 | 2012-10-11 | The Scripps Research Institute | Chromosomal landing pads and related uses |
Non-Patent Citations (6)
| Title |
|---|
| CHRISTINE LATTENMAYER ; MARTINA LOESCHEL ; WILLIBALD STEINFELLNER ; EVELYN TRUMMER ; DETHARDT MUELLER ; KORNELIA SCHRIEBL ; KAROLA: "Identification of transgene integration loci of different highly expressing recombinant CHO cell lines by FISH", CYTOTECHNOLOGY, KLUWER ACADEMIC PUBLISHERS, DO, vol. 51, no. 3, 15 November 2006 (2006-11-15), Do, pages 171 - 182, XP019448503, ISSN: 1573-0778, DOI: 10.1007/s10616-006-9029-0 * |
| HONG ZHOU, ZHI-GANG LIU, ZHI-WEI SUN, YING HUANG, WEI-YUAN YU: "Generation of stable cell lines by site-specific integration of transgenes into engineered Chinese hamster ovary strains using an FLP-FRT system.", JOURNAL OF BIOTECHNOLOGY, ELSEVIER, AMSTERDAM NL, vol. 147, no. 2, 17 May 2010 (2010-05-17), Amsterdam NL, pages 122 - 129, XP002711613, ISSN: 0168-1656, DOI: 10.1016/J.JBIOTEC.2010.03.020 * |
| LEWIS N E, LIU X, LI Y, NAGARAJAN H, YERGANIAN G, O'BRIEN E, BORDBAR A, ROTH A M, ROSENBLOOM J, BIAN C, XIE M, CHEN W, LI N, BAYCI: "Genomic landscapes of Chinese hamster ovary cell lines as revealed by the Cricetulus griseus draft genome.", NATURE BIOTECHNOLOGY, vol. 31, no. 8, 1 August 2013 (2013-08-01), pages 759 - 765, XP002753785, ISSN: 1546-1696, DOI: 10.1038/nbt.2624 * |
| MICHEL SADELAIN, EIRINI P. PAPAPETROU, FREDERIC D. BUSHMAN: "Safe harbours for the integration of new DNA in the human genome", NATURE REVIEWS CANCER, NATURE PUB. GROUP, vol. 12, no. 1, 1 January 2011 (2011-01-01), pages 51 - 58, XP055018235, ISSN: 1474175X, DOI: 10.1038/nrc3179 * |
| SCOTT BAHR;LAURA CORTNER;SARA LADLEY;TRISSA BORGSCHULTE: "Evaluating the effect of chromosomal context on zinc finger nuclease efficiency", BMC PROCEEDINGS, BIOMED CENTRAL LTD, LONDON UK, vol. 7, no. Suppl 6, 4 December 2013 (2013-12-04), London UK, pages P3, XP021170326, ISSN: 1753-6561, DOI: 10.1186/1753-6561-7-S6-P3 * |
| XUN XU, HARISH NAGARAJAN, NATHAN E LEWIS, SHENGKAI PAN, ZHIMING CAI, XIN LIU, WENBIN CHEN, MIN XIE, WENLIANG WANG, STEPHANIE HAMMO: "The genomic sequence of the Chinese hamster ovary (CHO)-K1 cell line", NATURE BIOTECHNOLOGY, GALE GROUP INC., NEW YORK, US, vol. 29, no. 8, us, pages 735 - 741, XP055223996, ISSN: 1087-0156, DOI: 10.1038/nbt.1932 * |
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| US9816110B2 (en) | 2017-11-14 |
| SG11201703149RA (en) | 2018-02-27 |
| CN107109434A (zh) | 2017-08-29 |
| US20240018553A1 (en) | 2024-01-18 |
| US20210171984A1 (en) | 2021-06-10 |
| EA201790698A1 (ru) | 2018-01-31 |
| AU2015335921B2 (en) | 2020-06-25 |
| CA2965495A1 (en) | 2016-04-28 |
| BR112017008022A2 (pt) | 2018-02-20 |
| TWI780847B (zh) | 2022-10-11 |
| CA2965495C (en) | 2022-05-24 |
| AU2015335921A1 (en) | 2017-05-04 |
| MY184103A (en) | 2021-03-18 |
| JP2017535258A (ja) | 2017-11-30 |
| TWI747808B (zh) | 2021-12-01 |
| TW201629229A (zh) | 2016-08-16 |
| EP3209785B1 (en) | 2019-07-31 |
| TW202144576A (zh) | 2021-12-01 |
| US20160115502A1 (en) | 2016-04-28 |
| WO2016064999A1 (en) | 2016-04-28 |
| IL251674B (en) | 2020-10-29 |
| US11788102B2 (en) | 2023-10-17 |
| MX2017005306A (es) | 2018-01-09 |
| EP3209785A1 (en) | 2017-08-30 |
| JP6668340B2 (ja) | 2020-03-18 |
| US20200002732A1 (en) | 2020-01-02 |
| KR102243243B1 (ko) | 2021-04-22 |
| US20180030480A1 (en) | 2018-02-01 |
| IL251674A0 (en) | 2017-06-29 |
| US11268109B2 (en) | 2022-03-08 |
| KR20170096999A (ko) | 2017-08-25 |
| AR102420A1 (es) | 2017-03-01 |
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