植物学报 ›› 2018, Vol. 53 ›› Issue (4): 528-541.DOI: 10.11983/CBB18004

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CRISPR/Cas9基因组定点编辑中脱靶现象的研究进展

王影1,2,*(), 李相敢2, 邱丽娟1,*()   

  1. 1中国农业科学院作物科学研究所, 北京 100081
    2先正达生物科技(中国)有限公司, 北京 102206
  • 收稿日期:2018-01-03 接受日期:2018-03-29 出版日期:2018-07-01 发布日期:2018-09-11
  • 通讯作者: 王影,邱丽娟
  • 作者简介:† 共同第一作者。

Research Progress in Off-target in CRISPR/Cas9 Genome Editing

Wang Ying1,2,*(), Li Xianggan2, Qiu Lijuan1,*()   

  1. 1Institute of Crop Sciences, Chinese Academy of Agricultural Sciences, Beijing 100081, China
    2Syngenta Biotechnology China Co, LTD, Beijing 102206, China
  • Received:2018-01-03 Accepted:2018-03-29 Online:2018-07-01 Published:2018-09-11
  • Contact: Wang Ying,Qiu Lijuan
  • About author:† These authors contributed equally to this paper

摘要: 近年来, CRISPR定点编辑技术发展迅猛, 在动物、植物和微生物中均得到广泛应用。其中, 备受关注的脱靶现象也是研究的热点, 迄今已取得了重要进展。该文介绍了脱靶现象的产生原理及体内和体外检测脱靶现象的方法, 评价了通过改进sgRNA设计和优化CRISPR系统等来降低脱靶率的方法。在植物基因组定点编辑过程中, 应适时检测脱靶现象, 提高脱靶检测的精确度和准确度。

关键词: CRISPR/Cas9, 定点编辑, 脱靶

Abstract: CRISPR/Cas9 technology has rapidly developed in recent years. It is widely used for study of animals, plants and microorganisms. Among all developments, the use of the technology for the off-target issue has progressed, with important breakthroughs for this hot topic. This paper elucidates the principle of off-target generation and the method for detecting off target both in vitro and in vivo. Also, how to reduce off-targets by improving single-guide RNA design and optimizing the CRISPR system is evaluated. Further more, detecting off-targets at the right time and improving the precision and accuracy of off-target detection are proposed for plant site-specific genome editing.

Key words: CRISPR/Cas9, site specific editing, off-target