“藕断丝连”的CRISPR/Cas:基因编辑中靶点滞留的作用与挑战  

Target Residence of CRISPR/Cas in Genome Editing

作　　者：冯依力[1,2] 陈若丹 谢安勇 FENG Yi-Li;CHEN Ruo-Dan;XIE An-Yong(Department of General Surgery,Sir Run Run Shaw Hospital,Zhejiang University School of Medicine,Hangzhou 310019,China;Institute of Translational Medicine,Zhejiang University School of Medicine,Hangzhou 310029,China)

机构地区：[1]浙江大学医学院附属邵逸夫医院普外科,杭州310019 [2]浙江大学转化医学研究院,杭州310029

出　　处：《生物化学与生物物理进展》2024年第10期2621-2636,共16页Progress In Biochemistry and Biophysics

基　　金：国家自然科学基金(32371348,32071439)资助项目。

摘　　要：成簇规律间隔短回文重复(clustered regulation interspaced short palindromic repeats,CRISPR)和CRISPR相关蛋白质(CRISPR-associated protein,Cas)系统被广泛应用于基因组编辑、转录调控以及细胞实时成像等,并已在农业、工业和医学等领域展示出巨大的应用潜力。该技术的应用取决于CRISPR/Cas的五大属性:靶向、解旋、切割、滞留和旁切。本综述将主要以化脓链球菌(Streptococcus pyogenes)的CRISPR/Cas9为例,聚焦于CRISPR/Cas的滞留属性,梳理相关进展,讨论其在基因编辑技术开发中的应用与挑战。The clustered regularly interspaced short palindromic repeats(CRISPR)/CRISPR-associated protein 9(Cas9)is widely used for targeted genomic and epigenomic modifications,transcriptional regulation and real-time cell imaging,and has already demonstrated great potential for applications in agriculture,industry and medicine.The promise of the technology depends upon the five intrinsic properties of CRISPR/Cas:targeting,target unwinding,target cutting,target residence,and collateral cleavage.Here,mainly using Streptococcus pyogenes CRISPR/Cas9 as example,we will focus on the target residence of CRISPR/Cas in applications of the CRISPR/Cas technology,summarize the recent progress,and discuss the effect of CRISPR/Cas target binding and residence on DNA double strand break repair pathway choices and the opportunities that CRISPR/Cas target residence presents to optimize the CRISPR/Cas technology.

关 键 词：CRISPR/Cas9 靶点滞留 靶点解离 DNA双链修复途径选择 基因编辑异质性 

分 类 号：Q78[生物学—分子生物学]