公开(公告)号：EP3177718A2

公开(公告)日：2017-06-14

申请号：EP15830407.1

申请日：2015-07-30

申请人： President and Fellows of Harvard College

发明人： LIU, David, R. ,  DAVIS, Kevin

IPC分类号： C12N9/22 ,  C07K14/315 ,  C07K14/195

CPC分类号： C12N15/907 ,  C07K14/35 ,  C07K14/721 ,  C07K2319/92 ,  C12N9/0071 ,  C12N9/22 ,  C12N9/78 ,  C12N15/63 ,  C12P19/34 ,  C12Y114/11 ,  C12Y305/04

摘要： Some aspects of this disclosure provide compositions, methods, systems, and kits for controlling the activity of RNA-programmable endonucleases, such as Cas9, or for controlling the activity of proteins comprising a Cas9 variant fused to a functional effector domain, such as a nuclease, nickase, recombinase, deaminase, transcriptional activator, transcriptional repressor, or epigenetic modifying domain. For example, the inventive proteins provided comprise a ligand-dependent intein, the presence of which inhibits one or more activities of the protein (e.g., gRNA binding, enzymatic activity, target DNA binding). The binding of a ligand to the intein results in self-excision of the intein, restoring the activity of the protein.

摘要翻译： 本公开的一些方面提供了用于控制RNA可编程核酸内切酶例如Cas9的活性或用于控制包含与功能性效应子结构域融合的Cas9变体的蛋白质的活性的组合物，方法，系统和试剂盒，所述功能性效应器域例如核酸酶 ，切口酶，重组酶，脱氨酶，转录激活物，转录阻遏物或表观遗传修饰结构域。 例如，提供的本发明蛋白质包含配体依赖性内含肽，其存在抑制蛋白质的一种或多种活性（例如，gRNA结合，酶促活性，靶DNA结合）。 配体与内含肽的结合导致内含肽的自我切除，恢复蛋白质的活性。