CRISPR-Cas9介导的基因组编辑技术的研究进展  

CRISPR-Cas9-based Genome Engineering

作　　者：郑小梅[1,2] 张晓立[1,3] 于建东[1,2] 郑平[1,2] 孙际宾[1,2] 

机构地区：[1]中国科学院系统微生物工程重点实验室,天津300308 [2]中国科学院天津工业生物技术研究所,天津300308 [3]天津科技大学生物工程学院,天津300457

出　　处：《生物技术进展》2015年第1期1-9,I0003,F0003,共11页Current Biotechnology

基　　金：国家863计划项目(2013AA020302);国家自然科学基金面上项目(31370113)资助

摘　　要：CRISPR-Cas(clustered regularly interspaced short palindromic repeats-CRISPR-associated proteins)系统为细菌与古生菌中抵御外源病毒或质粒DNA入侵的获得性免疫系统。该系统在crRNA的指导下,使核酸酶Cas识别并降解外源DNA。其中,Ⅱ型CRISPR-Cas系统最为简单,仅包括一个核酸酶Cas9与tracrRNA:crRNA二聚体便可完成其生物功能。基于CRISPR-Cas9的基因组编辑技术的核心为将tracrRNA:crRNA设计为引导RNA,在引导RNA的指导下Cas9定位于特定DNA序列上,进行DNA双链切割,实现基因组的定向编辑。CRISPR-Cas9系统以设计操纵简便、编辑高效与通用性广等优势成为新一代基因组编辑技术,为基因组定向改造调控与应用等带来突破性革命。从CRISPR-Cas9介导的基因组编辑技术的发展与应用等方面综述其最新研究进展,并着重介绍该技术的关键影响因素,为相关研究者提供参考。CRISPR-Cas （clustered regularly interspaced short palindromic repeats-CRISPR-associated proteins） systems are the adaptive immune system in bacteria and arehaea that defends against infectious viruses and plasmids. Immunity is mediated by Cas nucleases, which use small RNA guides （the crRNAs） to specify a cleavage site within the genome of invading nucleic acids. In type Ⅱ CRISPR-Cas systems, the DNA-eleaving activity is performed by a single enzyme Cas9 guided by an RNA duplex. Using synthetic single RNA guides, Cas9 can be reprogrammed to create specific double-stranded DNA breaks in the genomes of a variety of organisms, ranging from human cells to bacteria, and thus constitutes a powerful tool for genetic engineering. In this review, we described the development and applications of CRISPR-Cas9 system with highlighting the practical considerations for implementing CRISPR-Cas9 technology.

关 键 词：CRISPR-Cas系统 Cas9 基因组编辑技术 

分 类 号：Q811.4[生物学—生物工程]