CRISPR/Cas9系统中的脱靶效应及检测技术研究进展  

Research Progress on Off-target Effects and Detection Techniques in CRISPR/Cas9 Systems

作　　者：张晨[1] 雷展 李凯 商颖[1] 许文涛 ZHANG Chen;LEI Zhan;LI Kai;SHANG Ying;XU Wen-tao(College of Agriculture and Food,Kunming University of Science and Technology,Kunming 650504;Beijing Advanced Innovation Center for Food Nutrition and Human Health,College of Food Science and Nutritional Engineering,China Agricultural University,Beijing 100083;Key Laboratory of Safety Assessment of Genetically Modified Organism(Food Safety),Ministry of Agriculture,Beijing 100083)

机构地区：[1]昆明理工大学农业与食品学院,昆明650504 [2]中国农业大学食品科学与营养工程学院北京食品营养与人类健康高精尖创新中心,北京100083 [3]农业部农业转基因生物安全评价(食用)重点实验室,北京100083

出　　处：《生物技术通报》2020年第3期78-87,共10页Biotechnology Bulletin

基　　金：转基因重大专项(2018ZX08011-06B)

摘　　要：聚类规则间隔的短回文重复序列(Clustered regularly interspaced short palindromic repeats,CRISPR)及其相关蛋白(Cas9蛋白)已经成为精确编辑基因组的强大工具。CRISPR/Cas9系统源自于细菌和古生菌的免疫机制,结构简单、易于设计,仅需改变单引导RNA(single-guide RNA,sgRNA)上的一小段序列即可快速实现对不同基因位点的编辑。与锌指核酸酶(Zinc finger nucleases,ZFN)和转录激活因子样效应物核酸酶(Transcription activator-like effector nucleases,TALEN)技术相比,CRISPR/Cas9系统基因编辑效率更高、特异性更强。但在应用过程中不可避免的脱靶事件严重限制了CRISPR/Cas9系统的进一步发展,因此本篇文章就CRISPR/Cas9系统的脱靶类型、影响因素、降低策略以及脱靶检测技术进行综述。Clustered regularly interspaced short palindromic repeats(CRISPR)and its associated protein Cas9 have become as a powerful tool for precise genome editing.The CRISPR/Cas9 system,derived from the immune mechanism of bacteria and archaea,is simple in structure and easy to design.Different gene site can be quickly edited by simply changing a small sequence on the sgRNA.Compared with the zinc finger nucleases and transcription activator-like effector nucleases,the gene editing efficiency by CRISPR/Cas9 system is higher and the specificity is stronger.However,the inevitable off-target event in the application process severely limits the further development of the CRISPR/Cas9 system.Therefore,this article reviews the off-target type,influencing factors,reduction strategies and off-target detection technologies in applying the CRISPR/Cas9 system.

关 键 词：CRISPR/Cas9 脱靶效应 影响因素 降低策略 检测技术 

分 类 号：Q78[生物学—分子生物学]