CRISPR-Cas9-based Genome Engineering

doi:10.3969/j.issn.2095-2341.2015.01.01

Curr. Biotech. ›› 2015, Vol. 5 ›› Issue (1): 1-9.DOI: 10.3969/j.issn.2095-2341.2015.01.01

CRISPR-Cas9-based Genome Engineering

ZHENG Xiao-mei, ZHANG Xiao-li, YU Jian-dong , ZHENG Ping, , SUN Ji-bin,

1.Key Laboratory of Systems Microbial Biotechnology, Chinese Academy of Sciences, Tianjin 300308, China；

2.Tianjin Institute of Industrial Biotechnology, Chinese Academy of Sciences, Tianjin 300308, China；

3.School of Biological Engineering, Tianjin University of Science and Technology, Tianjin 300457, China

Received:2014-11-04 Online:2015-01-25 Published:2014-12-04

CRISPR-Cas9介导的基因组编辑技术的研究进展

郑小梅1,2,张晓立1,3,于建东1,2,郑平1,2,孙际宾1,2*

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

通讯作者: 孙际宾，研究员，博士生导师，主要从事微生物代谢工程与系统生物技术研究。Tel：022-84861949；E-mail：sun_jb@tib.cas.cn
作者简介:郑小梅，助理研究员，博士，从事黑曲霉分子生物学与系统生物学研究。E-mail：zheng_xm@tib.cas.cn。
基金资助:
国家863计划项目(2013AA020302)；国家自然科学基金面上项目(31370113)资助。

Abstract

Abstract: CRISPR-Cas (clustered regularly interspaced short palindromic repeats-CRISPR-associated proteins) systems are the adaptive immune system in bacteria and archaea 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-cleaving 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.

Key words: CRISPR-Cas system, Cas9, genome engineering

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。

关键词: CRISPR-Cas系统, Cas9, 基因组编辑技术

ZHENG Xiao-mei1,2, ZHANG Xiao-li1,3, YU Jian-dong 1,2, ZHENG Ping1,2,SUN Ji-bin1,2*. CRISPR-Cas9-based Genome Engineering[J]. Curr. Biotech., 2015, 5(1): 1-9.

郑小梅,张晓立,于建东,郑平,孙际宾,. CRISPR-Cas9介导的基因组编辑技术的研究进展[J]. 生物技术进展, 2015, 5(1): 1-9.

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CRISPR-Cas9-based Genome Engineering

CRISPR-Cas9介导的基因组编辑技术的研究进展

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