新一代精准基因编辑工具CRISPR/Cas9的技术优势与应用局限  

The Advantages and Limitations of CRISPR/Cas9-based Gene Editing Technology

作　　者：吴言[1] 郝雅荞 韦璇[1,2] 沈琦 柳叶飞 王升厚[3] 赵洪新[1] WU Yan;HAO Ya-qiao;WEI Xuan;SHEN Qi;LIU Ye-fei;WANG Sheng-hou;ZHAO Hong-xin(Zhefiang Province Key Laboratory of Plant Secondary Metabolism and Regulation, College of Life Sciences, Zhejiang Sci-Tech University, Hangzhou 310018;College of Life Science, Shenyang Normal University, Shenyang 110034;Experimental Teaching Center, Shenyang Normal University, Shenyang 110034)

机构地区：[1]浙江理工大学生命科学学院,杭州310018 [2]沈阳师范大学生命科学学院,沈阳110034 [3]沈阳师范大学教学实验中心,沈阳110034

出　　处：《生物技术通报》2018年第5期1-8,共8页Biotechnology Bulletin

基　　金：浙江省科技厅公益技术应用研究计划(2014003020);浙江理工大学人才基金项目(11612932611513)

摘　　要：CRISPR/Cas9(Clustered regularly interspaced short palindromic repeats/Cas9)是继锌指核酸酶(ZFNs)和类转录激活因子效应物核酸酶(TALENs)基因编辑技术之后的第三代基因编辑技术。CRISPR/Cas9在细菌和古生菌中广泛存在,是细菌在长期进化过程中形成的一种"适应性免疫防御",能够针对噬菌体感染、质粒接合和转化所造成的外源导入基因特异性识别、降解入侵的外源DNA,CRISPR/Cas9通过一段20 bp的短RNA来识别打靶位点的精准编辑技术。CRISPR/Cas9具有设计操作简便、编辑高效和通用性广等优点,是新一代具有革命意义的精准基因编辑技术。从CRISPR/Cas9的发现、作用机理、基因编辑以及应用局限等方面进行归纳总结,旨为理解其工作原理和精准基因编辑技术应用提供参考。CRISPR/Cas9 is becoming a hot spot in genome editing,and it provides a more efficient way for gene targeting than zinc finger endonuclease（ZFN）and transcription activator-like effector nuclease（TALEN）. CRISPR/Cas9 system exists widely in bacteria andarchaea,carrying out the adaptive immune responses specifically recognizing the invaded genetic elements from phage infection,plasmidconjugation and transformation as well as and degrading exogenous DNA. CRISPR/Cas9 is a precise editing technology by which the target sitesare recognized by a segment of 20 bp small RNA. CRISPR/Cas9 has the advantages of simple design and operation,high efficiency,and broadgenerality,thus it is a novel and revolutionary precise gene editing technology. This review induced and summarized the aspects of the discoveryof CRISPR/Cas,mechanisms,gene editing,and application limitations,aiming at providing references for understanding its mechanisms and applying precise gene editing technology.

关 键 词：CRISPR/Cas9 基因编辑 脱靶效应 sgRNA 

分 类 号：Q78[生物学—分子生物学]