TWI780847B - 新穎之ｃｈｏ整合位點及其用途 - Google Patents
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|---|---|---|---|---|
| SG11201703149RA (en) | 2014-10-23 | 2018-02-27 | Regeneron Pharma | Novel cho integration sites and uses thereof |
| 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 |
| EP3445781A1 (en) | 2016-04-20 | 2019-02-27 | Regeneron Pharmaceuticals, Inc. | Compositions and methods for making antibodies based on use of expression-enhancing loci |
| EP3504328A1 (en) | 2016-08-24 | 2019-07-03 | Regeneron Pharmaceuticals, Inc. | Host cell protein modification |
| JP2020501593A (ja) * | 2016-12-20 | 2020-01-23 | ディベロップメント センター フォー バイオテクノロジーＤｅｖｅｌｏｐｍｅｎｔ Ｃｅｎｔｅｒ Ｆｏｒ Ｂｉｏｔｅｃｈｎｏｌｏｇｙ | 改変細胞、調製方法、及び構築物 |
| WO2018148196A1 (en) * | 2017-02-07 | 2018-08-16 | Sigma-Aldrich Co. Llc | Stable targeted integration |
| JP7637470B2 (ja) * | 2017-02-17 | 2025-02-28 | ロンザ リミテッド | アデノ随伴ウイルスを生産するための哺乳動物細胞 |
| WO2018175153A1 (en) * | 2017-03-19 | 2018-09-27 | Applied Stemcell, Inc. | Novel 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 |
| US20200318106A1 (en) * | 2017-12-22 | 2020-10-08 | Bio-Rad Laboratories, Inc. | Controlling phenotype of organisms with crispr/cas gene targeting |
| IL275462B1 (en) * | 2017-12-22 | 2025-09-01 | Genentech Inc | Targeted integration of nucleic acids |
| 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 |
| 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 |
| 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 |
| 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 |
| 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 |
| AU2019403279A1 (en) * | 2018-12-21 | 2021-07-01 | Genentech, Inc. | Targeted integration of nucleic acids |
| SG11202110076PA (en) | 2019-04-02 | 2021-10-28 | Chugai Pharmaceutical Co Ltd | Method of introducing target-specific foreign gene |
| EP3990649A1 (en) | 2019-06-26 | 2022-05-04 | 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 |
| CN116134136A (zh) * | 2020-06-02 | 2023-05-16 | 康泰伦特药物解决方案有限责任公司 | 具有多个用于基因插入的Dock的细胞系 |
| CA3172453A1 (en) * | 2020-06-24 | 2021-12-30 | Amy Shen | Targeted integration of nucleic acids |
| EP4229205B1 (en) | 2020-10-15 | 2025-05-14 | F. Hoffmann-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 |
| US20230287460A1 (en) | 2021-10-18 | 2023-09-14 | Regeneron Pharmaceuticals, Inc. | Mammalian cells comprising integrated cas9 genes to produce stable integration sites, and mammalian cells comprising stable integration sites and other sites |
| KR20240099288A (ko) | 2021-10-18 | 2024-06-28 | 리제너론 파마슈티칼스 인코포레이티드 | 아데노바이러스-연관 바이러스 폴리뉴클레오티드를 포함하는 진핵세포 |
| TW202330926A (zh) | 2021-10-18 | 2023-08-01 | 美商再生元醫藥公司 | 聚核苷酸之受控制轉錄 |
| JP2025503925A (ja) * | 2022-01-28 | 2025-02-06 | ザ・トラスティーズ・オブ・コロンビア・ユニバーシティ・イン・ザ・シティ・オブ・ニューヨーク | 異種移植のための遺伝子改変 |
| 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 |
| TW202434888A (zh) | 2022-11-18 | 2024-09-01 | 美商再生元醫藥公司 | 用於偵測及測定包括生物流體在內之流體中之蛋白質結構及穩定性的方法 |
| 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 (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 |
| ATE539161T1 (de) | 1998-07-21 | 2012-01-15 | Millipore Corp | Ein ein allgegenwärtiges chromatin-öffnendes element (ucoe) enthaltendes polynucleotid |
| 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 |
| CA2745031C (en) * | 2008-12-04 | 2018-08-14 | Sangamo Biosciences, Inc. | Genome editing in rats using zinc-finger nucleases |
| CN105969807A (zh) | 2011-04-05 | 2016-09-28 | 斯克利普斯研究所 | 染色体着陆垫及相关用途 |
| EP3403672A1 (en) | 2011-04-20 | 2018-11-21 | Medlmmune, LLC | Antibodies and other molecules that bind b7-h1 and pd-1 |
| WO2012167192A2 (en) * | 2011-06-01 | 2012-12-06 | Precision Biosciences, Inc. | Methods and products for producing engineered mammalian cell lines with amplified transgenes |
| AU2012328682B2 (en) * | 2011-10-27 | 2017-09-21 | Sangamo Therapeutics, Inc. | Methods and compositions for modification of the HPRT locus |
| AU2013259647B2 (en) | 2012-05-07 | 2018-11-08 | Corteva Agriscience Llc | Methods and compositions for nuclease-mediated targeted integration of transgenes |
| SG11201703149RA (en) | 2014-10-23 | 2018-02-27 | Regeneron Pharma | Novel cho integration sites and uses thereof |
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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 (2)
| Title |
|---|
| 期刊 Lattenmayer C et al. "Identification of transgene integration loci of different highly expressing recombinant CHO cell lines by FISH" Cytotechnology vol.51 Springer 2006/11/15 p.171-182; * |
| 期刊 Xu X et al. "The genomic sequence of the Chinese hamster ovary (CHO)-K1 cell line" Nature biotechnology vol.29 no.8 Springer Nature Limited 2011/07/31 p.735-741 * |
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