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公开(公告)号：WO2016073433A1
公开(公告)日：2016-05-12
申请号：PCT/US2015/058760
申请日：2015-11-03
CPC分类号： C12N9/22 , C07K2319/09 , C07K2319/10 , C12N9/96 , C12N15/102 , C12N15/62 , C12N15/86 , C12N15/907 , C12N2310/3513 , C12N2750/14141
摘要： A composition is disclosed that comprises at least one protein component of an RNA-guided endonuclease (RGEN) and at least one cell-penetrating peptide (CPP), wherein the RGEN protein component and CPP are covalently or non-covalently linked to each other in an RGEN protein-CPP complex. The RGEN protein-CPP complex can traverse (i) a cell membrane, or (ii) a cell wall and cell membrane, of a cell. The RGEN protein component of an RGEN protein-CPP complex in certain embodiments can be associated with a suitable RNA component to provide an RGEN capable of specific DNA targeting. Further disclosed are compositions comprising at least one protein component of a guide polynucleotide/Cas endonuclease complex and at least one CPP, as well as methods of delivering RGEN proteins into microbial cells, as well as methods of targeting DNA with RGENs.
摘要翻译： 公开了包含RNA引导的内切核酸酶（RGEN）和至少一种细胞穿透肽（CPP）的至少一种蛋白质组分的组合物，其中所述RGEN蛋白质组分和CPP彼此共价或非共价连接 RGEN蛋白-CPP复合物。 RGEN蛋白-CPP复合物可以穿过（i）细胞膜，或（ii）细胞的细胞壁和细胞膜。 在一些实施方案中，RGEN蛋白-CPP复合物的RGEN蛋白组分可以与合适的RNA组分相关联，以提供能够进行特异性DNA靶向的RGEN。 还公开了包含导向多核苷酸/ Cas内切核酸酶复合物和至少一种CPP的至少一种蛋白质组分的组合物，以及将RGEN蛋白递送到微生物细胞中的方法，以及用RGEN靶向DNA的方法。
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2.发明申请PROTECTED DNA TEMPLATES FOR GENE MODIFICATION AND INCREASED HOMOLOGOUS RECOMBINATION IN CELLS AND METHODS OF USE 审中-公开用于基因修饰的受保护的DNA模板和增加的细胞中的同源重组以及使用方法
公开(公告)号：WO2017066175A1
公开(公告)日：2017-04-20
申请号：PCT/US2016/056404
申请日：2016-10-11
发明人： FRISCH, Ryan L.
IPC分类号： C12N15/11
CPC分类号： C12N15/111 , C12N15/102 , C12N15/815 , C12N15/902 , C12N2310/20
摘要： Compositions and methods are provided for modifying a nucleotide sequence in the genome of a cell. The methods and compositions employ a guide polynucleotide, a protected polynucleotide modification template and a Cas endonuclease to modify a nucleotide sequence and/or to increase the frequency of homologous directed repair. The methods can further be used to decrease the frequency of off-site integration of any modification template. The present disclosure also describes methods for selecting a cell comprising a modified target site in its genome and methods for selecting a cell comprising a polynucleotide of interest inserted into a target site in its genome.
摘要翻译： 提供了用于修饰细胞基因组中的核苷酸序列的组合物和方法。 该方法和组合物使用引导多核苷酸，保护的多核苷酸修饰模板和Cas核酸内切酶修饰核苷酸序列和/或增加同源定向修复的频率。 该方法可以进一步用于降低任何修改模板的场外整合的频率。 本公开还描述了用于选择在其基因组中包含修饰的靶位点的细胞的方法以及用于选择包含插入其基因组中的靶位点中的目的多核苷酸的细胞的方法。
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