CRISPR/Cas9系统中sgRNA设计与脱靶效应评估

doi:10.16288/j.yczz.15-093

遗传 ›› 2015, Vol. 37 ›› Issue (11): 1125-1136.doi: 10.16288/j.yczz.15-093

CRISPR/Cas9系统中sgRNA设计与脱靶效应评估

谢胜松¹,张懿²,张利生¹,李广磊¹,赵长志¹,倪攀¹,赵书红¹

1. 华中农业大学,农业动物遗传育种与繁殖教育部重点实验室,武汉 430070;
2. 中国人民解放军第161医院妇产科,武汉 430010

收稿日期:2015-03-02 修回日期:2015-06-18 出版日期:2015-11-20 发布日期:2015-08-03
通讯作者: 赵书红,教授,博士生导师,研究方向：动物分子生物学与育种。E-mail: shzhao@mail.hzau.edu.cn E-mail:ssxie@mail.hzau.edu.cn
作者简介:谢胜松,副研究员,研究方向：动物功能基因组与疾病诊治。E-mail: ssxie@mail.hzau.edu.cn
基金资助:
中央高校基本科研业务费专项资金资助项目(编号：2662015BQ005),广东省分子与细胞工程育种团队(编号：2011A020102003)和国家自然科学基金项目(编号：31301226)资助

sgRNA design for the CRISPR/Cas9 system and evaluation of its off-target effects

Shengsong Xie¹,Yi Zhang²,Lisheng Zhang¹,Guanglei Li¹,Changzhi Zhao¹,Pan Ni¹,Shuhong Zhao¹

1. Key Lab of Agricultural Animal Genetics, Breeding, and Reproduction of Ministry of Education, Huazhong Agricultural University, Wuhan 430070, China;
2. Department of Obstetrics and Gynecology, No.161 Hospital of PLA, Wuhan 430010, China

Received:2015-03-02 Revised:2015-06-18 Online:2015-11-20 Published:2015-08-03

摘要/Abstract

摘要： 基于CRISPR/Cas9系统介导的第三代基因组编辑技术,已成功应用于动物、植物和微生物等诸多物种的基因组改造。如何提高CRISPR/Cas9技术的基因组编辑效率和最大限度降低脱靶风险一直是本领域的研究热点,而使用高效且特异的sgRNA(Small guide RNA)是基因组改造成功的关键性因素之一。目前,已有多款针对CRISPR/Cas9技术的sgRNA设计和/或脱靶效应评估软件,但不同的软件各有优缺点。本文重点对16款sgRNA 设计和脱靶效应评估在线和单机版软件的特点进行了阐述,通过制定38项评估指标对不同软件进行了比较分析,最后对11种用于检测基因组编辑效率和脱靶的实验方法,以及如何筛选高效且特异的sgRNA进行了归纳总结。

关键词: CRISPR/Cas9系统, 基因组编辑, sgRNA, 脱靶效应

Abstract: The third generation of CRISPR/Cas9-mediated genome editing technology has been successfully applied to genome modification of various species including animals, plants and microorganisms. How to improve the efficiency of CRISPR/Cas9 genome editing and reduce its off-target effects has been extensively explored in this field. Using sgRNA (Small guide RNA) with high efficiency and specificity is one of the critical factors for successful genome editing. Several software have been developed for sgRNA design and/or off-target evaluation, which have advantages and disadvantages respectively. In this review, we summarize characters of 16 kinds online and standalone software for sgRNA design and/or off-target evaluation and conduct a comparative analysis of these different kinds of software through developing 38 evaluation indexes. We also summarize 11 experimental approaches for testing genome editing efficiency and off-target effects as well as how to screen highly efficient and specific sgRNA.

Key words: CRISPR/Cas9 system, genome editing, sgRNA, off-target effects

谢胜松,张懿,张利生,李广磊,赵长志,倪攀,赵书红. CRISPR/Cas9系统中sgRNA设计与脱靶效应评估[J]. 遗传, 2015, 37(11): 1125-1136.

Shengsong Xie,Yi Zhang,Lisheng Zhang,Guanglei Li,Changzhi Zhao,Pan Ni,Shuhong Zhao. sgRNA design for the CRISPR/Cas9 system and evaluation of its off-target effects[J]. HEREDITAS(Beijing), 2015, 37(11): 1125-1136.

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CRISPR/Cas9系统中sgRNA设计与脱靶效应评估

sgRNA design for the CRISPR/Cas9 system and evaluation of its off-target effects

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