CRISPR/Cas9系统研究进展

doi:10.13523/j.cb.20171012

中国生物工程杂志

2017, Vol. 37

Issue (10): 86-92 DOI: 10.13523/j.cb.20171012

综述

CRISPR/Cas9系统研究进展

栗晓飞, 曹英秀, 宋浩

天津大学化工学院系统生物工程教育部重点实验室天津化学化工协同创新中心合成生物学平台天津 300072

CRISPR/Cas9 System:A Recent Progress

LI Xiao-fei, CAO Ying-xiu, SONG Hao

School of Chemical Engineering and Technology, Key Laboratory of Systems Bioengineering(Ministry of Education), SynBio Research Platform, Collaborative Innovation Center of Chemical Science and Engineering(Tianjin), Tianjin University, Tianjin 300072, China

全文: PDF(557 KB) HTML

摘要： CRISPR/Cas9系统的发展彻底改变了人们编辑DNA序列和调控目标基因表达水平的能力，从而为生物体的精确基因组编辑提供了有力的工具。简化后的CRISPR/Cas9系统由两部分组成：Cas9蛋白和sgRNA。其作用原理为sgRNA通过自身的Cas9把手与Cas9蛋白形成Cas9-sgRNA复合体，Cas9-sgRNA复合体中sgRNA的碱基互补配对区序列与目标基因的靶序列通过碱基互补配对原则进行配对结合，Cas9利用自身的核酸内切酶活性对目标DNA序列进行切割。与传统的基因组编辑技术相比，CRISPR/Cas9系统具有几大明显的优势：易用性、简便性、低成本、可编程性以及可同时编辑多个基因。CRISPR/Cas9基因组编辑技术以及衍生出来的CRISPRi和CRISPRa基因表达调控技术已经广泛应用于多种真核和原核生物中。综述了CRISPR/Cas9系统的起源、作用机理、在生物体中的应用和其衍生出的技术，并概述了其脱靶效应和未来前景。

关键词： CRISPR/Cas9; CRISPRi; CRISPRa; 基因表达调控; 基因组编辑

Abstract: The development of CRISPR/Cas9 system has revolutionized our ability to edit DNA and to modulate expression levels of target genes, thus providing powerful tools to accelerate the precise genome engineering of a wide range of organisms. The developed CRISPR/Cas9 system consists of the Cas9 protein and a programmed sgRNA. The Cas9 protein binds to the Cas9 handle of sgRNA and forms a Cas9-sgRNA complex. Then, the Cas9-sgRNA complex binds to specific DNA targets by Watson-Crick base pairing between the sgRNA and the DNA target, and the DNA will be cleaved due to the nuclease activity of the Cas9 protein. Compared with the traditional genome editing technologies, CRISPR/Cas9 system has several obvious advantages, inculding ease of use, simplicity, low cost, programmed and multiple genes editing. CRISPR/Cas9 genome editing technology and the derived CRISPRi and CRISPRa gene expression regulation techniques have been widely used in a variety of eukaryotic and prokaryotic organisms. Here, the origin and mechanism of CRISPR/Cas9 system, its application in organisms and its derived technology were reviewed, and its off-target effect and future prospects were outlined.

Key words: CRISPR/Cas9 CRISPRi CRISPRa Genome editing Gene expression regulation

收稿日期: 2017-05-12 出版日期: 2017-10-25

ZTFLH:

Q789

基金资助: 国家自然科学基金（21376174）、天津市自然科学基金（13JCYBJC40700）资助项目

通讯作者: 宋浩,hsong@tju.edu.cn E-mail: hsong@tju.edu.cn

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引用本文:

栗晓飞, 曹英秀, 宋浩. CRISPR/Cas9系统研究进展[J]. 中国生物工程杂志, 2017, 37(10): 86-92.

LI Xiao-fei, CAO Ying-xiu, SONG Hao. CRISPR/Cas9 System:A Recent Progress. China Biotechnology, 2017, 37(10): 86-92.

链接本文:

https://manu60.magtech.com.cn/biotech/CN/10.13523/j.cb.20171012 或 https://manu60.magtech.com.cn/biotech/CN/Y2017/V37/I10/86

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