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1. 发明申请

WO2017049240A1 SELECTION OF PLURIPOTENT CELLS FOR PRODUCTION OF FERTILE XY FEMALE MICE 审中-公开
标题翻译：选择用于生产可食用X射线母鼠的小鼠细胞
公开(公告)号：WO2017049240A1
公开(公告)日：2017-03-23
申请号：PCT/US2016/052345
申请日：2016-09-16
申请人： REGENERON PHARMACEUTICALS, INC.
发明人： SCHMAHL, Jennifer , FRENDEWEY, David , KUNO, Junko , SIAO, Chia-Jen , DROGUETT, Gustavo , BAI, Yu , AUERBACH, Wojtek
IPC分类号： A01K67/027 , C12N5/0735
CPC分类号： A01K67/0271 , A01K2207/12 , A01K2207/35 , A01K2227/105 , A01K2267/02 , C12N5/0606 , C12Q1/6876 , C12Q2600/136 , C12Q2600/156 , C12Q2600/158 , G01N33/5073
摘要： Methods and compositions are provided for generating F0 fertile XY female animals. The methods and compositions involve making XY pluripotent or totipotent animal cells, in vitro cell cultures, or embryos that are capable of producing a fertile female XY animal in an F0 generation. Such cells, embryos, and animals can be made by silencing a region of the Y chromosome. Optionally, the cells can also be cultured in feminizing medium such as a low-osmolality medium and/or can be modified to decrease the level and/or activity of an Sry protein. Methods and compositions are also provided for silencing a region of the Y chromosome in an XY pluripotent or totipotent animal cell, or in vitro cell cultures, embryos, or animals derived therefrom, by maintaining an XY pluripotent or totipotent animal cell in a feminizing medium. Methods and compositions are also provided for maintaining a population of XY pluripotent or totipotent animal cells in a feminizing medium and selecting cells or clones having increased capabilities for producing a fertile female XY animal in an F0 generation. Methods and compositions are also provided for screening for compounds with feminizing activity or for optimizing concentrations of components in feminizing media.
摘要翻译：提供了用于产生F0可育XY雌性动物的方法和组合物。所述方法和组合物包括制备XY多能或全能动物细胞，体外细胞培养物或能够在F0代中产生可育雌性XY动物的胚胎。可以通过沉默Y染色体的区域来制备这样的细胞，胚胎和动物。任选地，细胞还可以在女性化培养基如低渗透浓度培养基中培养和/或可被修饰以降低Sry蛋白的水平和/或活性。还提供了通过将XY多能或全能动物细胞维持在女性化培养基中，从而在XY多能或全能动物细胞或体外细胞培养物，胚胎或由其衍生的动物中沉默Y染色体区域的方法和组合物。还提供了用于在女性化培养基中维持XY多能或全能动物细胞群的方法和组合物，并选择具有增加在F0代中产生可育雌性XY动物的能力的细胞或克隆。还提供了用于筛选具有女性化活性的化合物或优化女性化培养基中组分浓度的方法和组合物。

2. 发明申请

WO2016061374A1 METHODS AND COMPOSITIONS FOR GENERATING OR MAINTAINING PLURIPOTENT CELLS 审中-公开
标题翻译：用于产生或维持PLUIPOTENT细胞的方法和组合物
公开(公告)号：WO2016061374A1
公开(公告)日：2016-04-21
申请号：PCT/US2015/055776
申请日：2015-10-15
申请人： REGENERON PHARMACEUTICALS, INC.
发明人： KUNO, Junko , AUERBACH, Wojtek , VALENZUELA, David M.
IPC分类号： C12N5/074 , C12N5/00
CPC分类号： C12N5/0696 , C12N2500/32 , C12N2500/38 , C12N2500/44 , C12N2500/60 , C12N2501/235 , C12N2501/727 , C12N2509/00
摘要： Methods and compositions are provided for generating or maintaining human iPS cells in culture. Methods include the use of a low osmolality medium to make human iPS cells, or use of a low osmolality medium to maintain human iPS cells. Methods for making targeted genetic modification to human iPS cells cultured in low osmolality medium are also included. Compositions include human iPS cells cultured and maintained using the low osmolality medium defined herein.
摘要翻译：提供了用于在培养物中产生或维持人iPS细胞的方法和组合物。方法包括使用低渗透浓度培养基制备人iPS细胞，或使用低渗透浓度培养基来维持人类iPS细胞。还包括对在低渗透浓度培养基中培养的人iPS细胞进行靶向遗传修饰的方法。组合物包括使用本文定义的低渗透浓度培养基培养和维持的人iPS细胞。

3. 发明申请

WO2015200805A3 METHODS AND COMPOSITIONS FOR TARGETED GENETIC MODIFICATIONS AND METHODS OF USE 审中-公开
公开(公告)号：WO2015200805A3
公开(公告)日：2015-12-30
申请号：PCT/US2015/038001
申请日：2015-06-26
申请人： REGENERON PHARMACEUTICALS, INC.
发明人： FRENDEWEY, David , DROGUETT, Gustavo , GAGLIARDI, Anthony , KUNO, Junko , AUERBACH, Wojtek , VALENZUELA, David M.
IPC分类号： C12N5/0735 , C12N15/873 , C12N15/85 , C12N15/90
摘要： Methods and compositions are provided for generating targeted genetic modifications on the Y chromosome or a challenging target locus. Compositions include an in vitro culture comprising an XY pluripotent and/or totipotent animal cell (i.e., XY ES cells or XY iPS cells) having a modification that decreases the level and/or activity of an Sry protein; and, culturing these cells in a medium that promotes development of XY F0 fertile females. Such compositions find use in various methods for making a fertile female XY non-human mammal in an F0 generation.

4. 发明申请

WO2015088643A1 METHODS AND COMPOSITIONS FOR THE TARGETED MODIFICATION OF A GENOME 审中-公开
标题翻译：基因组目标修饰的方法和组合
公开(公告)号：WO2015088643A1
公开(公告)日：2015-06-18
申请号：PCT/US2014/060788
申请日：2014-10-15
申请人： REGENERON PHARMACEUTICALS, INC.
发明人： FRENDEWEY, David , AUERBACH, Wojtek , LAI, Ka-Man Venus , MUJICA, Alexander O. , LEE, Jeffrey D. , DROGUETT, Gustavo , TRZASKA, Sean , HUNT, Charleen , GAGLIARDI, Anthony , KUNO, Junko , VALENZUELA, David M. , YANCOPOULOS, George D.
IPC分类号： C12N15/85 , C12N9/22
CPC分类号： C12N15/8509 , A01K67/0276 , A01K67/0278 , A01K2217/072 , A01K2217/075 , A01K2227/105 , A01K2267/0362 , A01K2267/0387 , C12N9/22 , C12N15/1024 , C12N15/85 , C12N15/907 , C12N2015/8527 , C12N2800/40 , C12N2810/00 , C12N2810/10 , C12N2810/40 , C12N2999/007
摘要： Compositions and methods are provided for modifying a genomic locus of interest in a eukaryotic cell, a mammalian cell, a human cell or a non-human mammalian cell using a large targeting vector (LTVEC) comprising various endogenous or exogenous nucleic acid sequences as described herein. Further methods combine the use of the LTVEC with a CRISPR/Cas system. Compositions and methods for generating a genetically modified non-human animal comprising one or more targeted genetic modifications in their germline are also provided.
摘要翻译：提供组合物和方法用于使用包含本文所述的各种内源或外源核酸序列的大靶向载体（LTVEC）修饰真核细胞，哺乳动物细胞，人细胞或非人哺乳动物细胞中的感兴趣的基因组座位。。其他方法将LTVEC与CRISPR / Cas系统的使用相结合。还提供了用于产生遗传修饰的非人动物的组合物和方法，其包含其种系中的一种或多种靶向遗传修饰。

5. 发明申请

WO2020131632A1 NUCLEASE-MEDIATED REPEAT EXPANSION 审中-公开
公开(公告)号：WO2020131632A1
公开(公告)日：2020-06-25
申请号：PCT/US2019/066317
申请日：2019-12-13
申请人： REGENERON PHARMACEUTICALS, INC.
发明人： KAJIMURA, Daisuke , SHARMA-KANNING, Aarti , DUBOSE, Brittany , DROGUETT, Gustavo , SIAO, Chia-Jen , KUNO, Junko , FRENDEWEY, David , ZAMBROWICZ, Brian
IPC分类号： C12N15/10 , A01K67/027 , C12N15/11
摘要： Nuclease-mediated methods for expanding repeats already present at a genomic locus are provided. Non-human animal genomes, non-human animal cells, and non-human animals comprising a heterologous hexanucleotide repeat expansion sequence inserted at an endogenous C9orf72 locus and methods of making such non-human animal cells and non-human animals through nuclease-mediated repeat expansion are also provided. Methods of using the non-human animal cells or non-human animals to identify therapeutic candidates that may be used to prevent, delay or treat one or more neurodegenerative disorders associated with repeat expansion at the C9orf72 locus are also provided.

6. 发明申请

WO2017049240A8 SELECTION OF PLURIPOTENT CELLS FOR PRODUCTION OF FERTILE XY FEMALE MICE 审中-公开
公开(公告)号：WO2017049240A8
公开(公告)日：2017-03-23
申请号：PCT/US2016/052345
申请日：2016-09-16
申请人： REGENERON PHARMACEUTICALS, INC.
发明人： SCHMAHL, Jennifer , FRENDEWEY, David , KUNO, Junko , SIAO, Chia-Jen , DROGUETT, Gustavo , BAI, Yu , AUERBACH, Wojtek
IPC分类号： A01K67/027 , C12N5/0735
摘要： Methods and compositions are provided for generating F0 fertile XY female animals. The methods and compositions involve making XY pluripotent or totipotent animal cells, in vitro cell cultures, or embryos that are capable of producing a fertile female XY animal in an F0 generation. Such cells, embryos, and animals can be made by silencing a region of the Y chromosome. Optionally, the cells can also be cultured in feminizing medium such as a low-osmolality medium and/or can be modified to decrease the level and/or activity of an Sry protein. Methods and compositions are also provided for silencing a region of the Y chromosome in an XY pluripotent or totipotent animal cell, or in vitro cell cultures, embryos, or animals derived therefrom, by maintaining an XY pluripotent or totipotent animal cell in a feminizing medium. Methods and compositions are also provided for maintaining a population of XY pluripotent or totipotent animal cells in a feminizing medium and selecting cells or clones having increased capabilities for producing a fertile female XY animal in an F0 generation. Methods and compositions are also provided for screening for compounds with feminizing activity or for optimizing concentrations of components in feminizing media.

7. 发明申请

WO2015188109A1 METHODS AND COMPOSITIONS FOR MODIFYING A TARGETED LOCUS 审中-公开
标题翻译：用于修改目标位置的方法和组合
公开(公告)号：WO2015188109A1
公开(公告)日：2015-12-10
申请号：PCT/US2015/034503
申请日：2015-06-05
申请人： REGENERON PHARMACEUTICALS, INC.
发明人： AUERBACH, Wojtek , FRENDEWEY, David , DROGUETT, Gustavo , GAGLIARDI, Anthony , KUNO, Junko , VALENZUELA, David M.
IPC分类号： C12N15/90
CPC分类号： C12N15/907 , A01K67/0278 , A01K2217/072 , A01K2227/105 , A01K2267/0387 , C07K14/7051 , C12N15/64 , C12N2810/00
摘要： Methods and compositions are provided for modifying one or more target loci in a cell. Such methods comprise providing a cell comprising a first polynucleotide encoding a first selection marker operably linked to a first promoter active in the cell, wherein the first polynucleotide further comprises a first recognition site for a first nuclease agent. A first nuclease agent is introduced into a cell, wherein the first nuclease agent induces a nick or double-strand break at the first recognition site. Further introduced into the cell is a first targeting vector comprising a first insert polynucleotide flanked by a first and a second homology arm that correspond to a first and a second target site located in sufficient proximity to the first recognition site. At least one cell is then identified comprising in its genome the first insert polynucleotide integrated at the target locus.
摘要翻译：提供了用于修饰细胞中一个或多个靶基因座的方法和组合物。这样的方法包括提供包含编码第一选择标记的第一多核苷酸的细胞，所述第一选择标记可操作地连接到细胞中活性的第一启动子，其中所述第一多核苷酸还包含第一核酸酶试剂的第一识别位点。将第一核酸酶试剂引入细胞中，其中第一核酸酶试剂在第一识别位点诱导切口或双链断裂。进一步引入细胞中的是第一靶向载体，其包含侧翼于第一和第二同源性臂的第一插入核苷酸，所述第一和第二同源性臂对应于位于第一识别位点附近的第一和第二靶位点。然后鉴定至少一个细胞，其在其基因组中包含整合在靶标位点的第一插入多核苷酸。

8. 发明公开

EP3732291A1 NUCLEASE-MEDIATED REPEAT EXPANSION 审中-公开
公开(公告)号：EP3732291A1
公开(公告)日：2020-11-04
申请号：EP19836402.8
申请日：2019-12-13
申请人： Regeneron Pharmaceuticals, Inc.
发明人： KAJIMURA, Daisuke , SHARMA-KANNING, Aarti , DUBOSE, Brittany , DROGUETT, Gustavo , SIAO, Chia-Jen , KUNO, Junko , FRENDEWEY, David , ZAMBROWICZ, Brian
IPC分类号： C12N15/10 , A01K67/027 , C12N15/11

9. 发明公开

EP3349573A1 SELECTION OF PLURIPOTENT CELLS FOR PRODUCTION OF FERTILE XY FEMALE MICE 审中-公开
公开(公告)号：EP3349573A1
公开(公告)日：2018-07-25
申请号：EP16778528.6
申请日：2016-09-16
申请人： Regeneron Pharmaceuticals, Inc.
发明人： SCHMAHL, Jennifer , FRENDEWEY, David , KUNO, Junko , SIAO, Chia-Jen , DROGUETT, Gustavo , BAI, Yu , AUERBACH, Wojtek
IPC分类号： A01K67/027 , C12N5/0735
CPC分类号： A01K67/0271 , A01K2207/12 , A01K2207/35 , A01K2227/105 , A01K2267/02 , C12N5/0606 , C12Q1/6876 , C12Q2600/136 , C12Q2600/156 , C12Q2600/158 , G01N33/5073
摘要： Methods and compositions are provided for generating F0 fertile XY female animals. The methods and compositions involve making XY pluripotent or totipotent animal cells, in vitro cell cultures, or embryos that are capable of producing a fertile female XY animal in an F0 generation. Such cells, embryos, and animals can be made by silencing a region of the Y chromosome. Optionally, the cells can also be cultured in feminizing medium such as a low-osmolality medium and/or can be modified to decrease the level and/or activity of an Sry protein. Methods and compositions are also provided for silencing a region of the Y chromosome in an XY pluripotent or totipotent animal cell, or in vitro cell cultures, embryos, or animals derived therefrom, by maintaining an XY pluripotent or totipotent animal cell in a feminizing medium. Methods and compositions are also provided for maintaining a population of XY pluripotent or totipotent animal cells in a feminizing medium and selecting cells or clones having increased capabilities for producing a fertile female XY animal in an F0 generation. Methods and compositions are also provided for screening for compounds with feminizing activity or for optimizing concentrations of components in feminizing media.

10. 发明公开

EP3207124A1 METHODS AND COMPOSITIONS FOR GENERATING OR MAINTAINING PLURIPOTENT CELLS 有权
标题翻译：产生或维持多潜能细胞的方法和组合物
公开(公告)号：EP3207124A1
公开(公告)日：2017-08-23
申请号：EP15787382.9
申请日：2015-10-15
申请人： Regeneron Pharmaceuticals, Inc.
发明人： KUNO, Junko , AUERBACH, Wojtek , VALENZUELA, David M.
IPC分类号： C12N5/074 , C12N5/00
CPC分类号： C12N5/0696 , C12N2500/32 , C12N2500/38 , C12N2500/44 , C12N2500/60 , C12N2501/235 , C12N2501/727 , C12N2509/00
摘要： Methods and compositions are provided for generating or maintaining human iPS cells in culture. Methods include the use of a low osmolality medium to make human iPS cells, or use of a low osmolality medium to maintain human iPS cells. Methods for making targeted genetic modification to human iPS cells cultured in low osmolality medium are also included. Compositions include human iPS cells cultured and maintained using the low osmolality medium defined herein.
摘要翻译：提供了用于在培养中产生或维持人类iPS细胞的方法和组合物。方法包括使用低渗透压培养基制造人类iPS细胞，或使用低渗透压培养基维持人类iPS细胞。还包括对在低渗透压培养基中培养的人iPS细胞进行靶向遗传修饰的方法。组合物包括使用本文定义的低渗透压介质培养和维持的人iPS细胞。

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