TWI747808B - 新穎之ｃｈｏ整合位點及其用途 - Google Patents
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Families Citing this family (38)
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|---|---|---|---|---|
| CN107109434A (zh) | 2014-10-23 | 2017-08-29 | 瑞泽恩制药公司 | 新颖cho整合位点和其用途 |
| 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 |
| SG10202010156XA (en) | 2016-04-20 | 2020-11-27 | Regeneron Pharma | Compositions and methods for making antibodies based on use of an expression-enhancing locus |
| CN109195986B (zh) | 2016-04-20 | 2023-01-03 | 瑞泽恩制药公司 | 基于使用表达增强性基因座来制备抗体的组合物和方法 |
| WO2018039499A1 (en) | 2016-08-24 | 2018-03-01 | Regeneron Pharmaceuticals, Inc. | Host cell protein modification |
| EP4130280A3 (en) * | 2016-12-20 | 2023-04-19 | Development Center for Biotechnology | Targeted integration sites in chinese hamster ovary cell genome |
| US20210309988A1 (en) * | 2017-02-07 | 2021-10-07 | Sigma-Aldrich Co. Llc | Stable targeted integration |
| EP3583205A1 (en) * | 2017-02-17 | 2019-12-25 | Lonza Ltd | Mammalian cells for producing adeno-associated viruses |
| US11959093B2 (en) | 2017-03-19 | 2024-04-16 | Applied Stemcell, Inc. | Integration sites and uses thereof |
| AU2018315428B2 (en) | 2017-08-11 | 2024-11-14 | 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 |
| CN111566208A (zh) * | 2017-12-22 | 2020-08-21 | 生物辐射实验室股份有限公司 | 利用CRISPR/Cas基因靶向控制生物体表型 |
| 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 |
| 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 |
| 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 |
| 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 |
| 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 |
| 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 |
| TW202039852A (zh) * | 2018-12-21 | 2020-11-01 | 美商建南德克公司 | 核酸之靶向整合 |
| IL286630B2 (en) | 2019-04-02 | 2025-05-01 | Chugai Pharmaceutical Co Ltd | Method for introducing a target-specific foreign gene |
| WO2020264253A1 (en) | 2019-06-26 | 2020-12-30 | 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 |
| BR112022024625A2 (pt) * | 2020-06-02 | 2023-02-23 | Catalent Pharma Solutions Llc | Construtos de ácido nucleico para a fabricação de proteína |
| KR20230027043A (ko) * | 2020-06-24 | 2023-02-27 | 제넨테크, 인크. | 핵산의 표적화 통합 |
| KR20230085929A (ko) | 2020-10-15 | 2023-06-14 | 에프. 호프만-라 로슈 아게 | Va rna 전사를 위한 핵산 구조체 |
| 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 |
| US20230304062A1 (en) | 2021-10-18 | 2023-09-28 | Regeneron Pharmaceuticals, Inc. | Controlled transcription of polynucleotides |
| CA3235566A1 (en) | 2021-10-18 | 2023-04-27 | Michael Goren | Mammalian cells comprising integrated cas9 genes to produce stable integration sites, and mammalian cells comprising stable integration sites and other sites |
| MX2024004647A (es) | 2021-10-18 | 2024-05-02 | Regeneron Pharma | Celulas eucariotas que comprenden polinucleotidos de virus asociados a adenovirus. |
| MX2024009088A (es) * | 2022-01-28 | 2024-07-31 | Univ Columbia | Modificaciones geneticas para xenotrasplantes. |
| 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 |
| US20250163468A1 (en) | 2023-11-21 | 2025-05-22 | Regeneron Pharmaceuticals, Inc. | Covalently surface modified adeno-associated virus production by in vitro conjugation, and purification of covalently surface modified adeno-associated virus |
Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20130130372A1 (en) * | 2007-06-04 | 2013-05-23 | Regeneron Pharmaceuticals, Inc. | Enhanced expression and stability regions |
Family Cites Families (14)
| 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 |
| WO2002081632A2 (en) | 2001-04-04 | 2002-10-17 | The Government Of The United States Of America, As Represented By The Secretary Of The Department Of Health And Human Services | 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 |
| AU2009322964B2 (en) * | 2008-12-04 | 2014-10-09 | Sangamo Therapeutics, Inc. | Genome editing in rats using zinc-finger nucleases |
| CA2832095A1 (en) | 2011-04-05 | 2012-10-11 | The Scripps Research Institute | Chromosomal landing pads and related uses |
| EP2699264B1 (en) | 2011-04-20 | 2018-03-14 | Medlmmune, LLC | Antibodies and other molecules that bind b7-h1 and pd-1 |
| DK2714936T3 (en) * | 2011-06-01 | 2019-03-25 | Prec Biosciences Inc | METHODS AND PRODUCTS FOR PRODUCING MANIPULATED MAMMAL CELL LINES WITH AMPLIFIED TRANSGENES |
| JP6188703B2 (ja) * | 2011-10-27 | 2017-08-30 | サンガモ セラピューティクス， インコーポレイテッド | Ｈｐｒｔ遺伝子座を修飾するための方法および組成物 |
| CA2871524C (en) | 2012-05-07 | 2021-07-27 | Sangamo Biosciences, Inc. | Methods and compositions for nuclease-mediated targeted integration of transgenes |
| CN107109434A (zh) | 2014-10-23 | 2017-08-29 | 瑞泽恩制药公司 | 新颖cho整合位点和其用途 |
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Patent Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20130130372A1 (en) * | 2007-06-04 | 2013-05-23 | Regeneron Pharmaceuticals, Inc. | Enhanced expression and stability regions |
Non-Patent Citations (3)
| Title |
|---|
| Lattenmayer C et al., "Identification of transgene integration loci of different highly expressing recombinant CHO cell lines by FISH", Cytotechnology, vol.51, p.171-182, 2006/11/15 |
| Lattenmayer C et al., "Identification of transgene integration loci of different highly expressing recombinant CHO cell lines by FISH", Cytotechnology, vol.51, p.171-182, 2006/11/15; * |
| Xu X et al., "The genomic sequence of the Chinese hamster ovary (CHO)-K1 cell line", Nature biotechnology, vol.29, no.8, p.735-741, 2011/07/31 * |
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| US11788102B2 (en) | 2023-10-17 |
| KR20170096999A (ko) | 2017-08-25 |
| TWI780847B (zh) | 2022-10-11 |
| TW202144576A (zh) | 2021-12-01 |
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