遗传 ›› 2015, Vol. 37 ›› Issue (10): 1011-1020.doi: 10.16288/j.yczz.15-066

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CRISPR/Cas9基因组编辑技术及其在动物基因组定点修饰中的应用

周金伟1, 徐绮嫔1, 姚婧2, 余树民1, 曹随忠1   

  1. 1. 四川农业大学动物医学院,成都 611130;
    2. 中国科学院动物研究所,计划生育生殖生物学国家重点实验室,北京 100101
  • 收稿日期:2015-02-03 出版日期:2015-10-20 发布日期:2015-10-20
  • 通讯作者: 曹随忠,教授,研究方向:兽医产科学。E-mail: suizhongcao@126.com
  • 作者简介:周金伟,硕士,专业方向:大动物遗传与修饰。E-mail: zhoujinwei08@126.com
  • 基金资助:
    国家自然科学基金项目(编号:31272400,31172379)和四川农业大学学科建设双支计划项目(编号:3570806)资助

CRISPR/Cas9 genome editing technique and its application in site-directed genome modification of animals

Jinwei Zhou1, Qipin Xu1, Jing Yao2, Shumin Yu1, Suizhong Cao1   

  1. 1. College of Veterinary Medicine, Sichuan Agricultural University, Chengdu 611130, China;
    2. State Key Laboratory of Reproductive Biology, Institute of Zoology, Chinese Academy of Sciences, Beijing 100101, China
  • Received:2015-02-03 Online:2015-10-20 Published:2015-10-20

摘要: CRISPR/Cas系统是细菌和古生菌中抵抗外源病毒或质粒入侵的获得性免疫系统,利用CRISPR RNAs (crRNAs)引导Cas核酸酶沉默入侵的核酸。通过分子生物学改造使Ⅱ型CRISPR/Cas系统成为一种高效的基因组定点修饰技术,并且比锌指核酸酶(Zinc-finger nucleases, ZFNs)和TALE核酸酶(Transcription activator like effector nucleases, TALENs)结构更简单,更容易设计和应用。文章主要介绍了CRISPR/Cas9系统成为高效基因组定点修饰技术的发展历程、Ⅱ型CRISPR/Cas的工作原理和改造过程以及在动物基因组定点修饰的应用,剖析了该技术存在的问题和现有改进方案,并与成功案例相结合展望了CRISPR/Cas9系统的应用前景,以期为动物性状改良和人类疾病动物模型的创立提供新思路。

关键词: CRISPR/Cas9, 基因组编辑技术, 人类疾病动物模型, 基因组定点修饰

Abstract: CRISPR/Cas system, which uses CRISPR RNAs (crRNAs) to guide Cas nuclease to silence invading nucleic acids, is self-defense system against exogenous virus or plasmid in bacteria and archaea. Through molecular modification, the typeⅡCRISPR/Cas system has become a highly efficient site-directed genome editing technique, which is simpler than zinc-finger nucleases (ZFNs) and transcription activator like effector nucleases (TALENs) and easier to be designed and applied. In this review, we summarize the evolutionary history of CRISPR/Cas9 system, the working principle and modification process of type Ⅱ CRISPR/Cas and its application in animal genome modification. We also analyze the existing problems and improvement program of the CRISPR/Cas9 system as well as its application prospect combined with successful cases, which may provide innovative perspectives on improving animal traits and establishing animal models of human diseases.

Key words: CRISPR/Cas9, genome editing technique, animal models of human disease, site-directed genome modification