CRISPR/Cas9系统中的脱靶效应及检测技术研究进展

doi:10.13560/j.cnki.biotech.bull.1985.2019-0871

摘要/Abstract

摘要： 聚类规则间隔的短回文重复序列（Clustered regularly interspaced short palindromic repeats,CRISPR）及其相关蛋白（Cas9蛋白）已经成为精确编辑基因组的强大工具。CRISPR/Cas9系统源自于细菌和古生菌的免疫机制,结构简单、易于设计,仅需改变单引导RNA（single-guide RNA,sgRNA）上的一小段序列即可快速实现对不同基因位点的编辑。与锌指核酸酶（Zinc finger nucleases,ZFN）和转录激活因子样效应物核酸酶（Transcription activator-like effector nucleases,TALEN）技术相比,CRISPR/Cas9系统基因编辑效率更高、特异性更强。但在应用过程中不可避免的脱靶事件严重限制了CRISPR/Cas9系统的进一步发展,因此本篇文章就CRISPR/Cas9系统的脱靶类型、影响因素、降低策略以及脱靶检测技术进行综述。

关键词: CRISPR/Cas9, 脱靶效应, 影响因素, 降低策略, 检测技术

Abstract: Clustered regularly interspaced short palindromic repeats（CRISPR）and its associated protein Cas9 have become as a powerful tool for precise genome editing. The CRISPR/Cas9 system,derived from the immune mechanism of bacteria and archaea,is simple in structure and easy to design. Different gene site can be quickly edited by simply changing a small sequence on the sgRNA. Compared with the zinc finger nucleases and transcription activator-like effector nucleases,the gene editing efficiency by CRISPR/Cas9 system is higher and the specificity is stronger. However,the inevitable off-target event in the application process severely limits the further development of the CRISPR/Cas9 system. Therefore,this article reviews the off-target type,influencing factors,reduction strategies and off-target detection technologies in applying the CRISPR/Cas9 system.

Key words: CRISPR/Cas9, off-target effect, influencing factors, reduction strategy, detection technology

张晨, 雷展, 李凯, 商颖, 许文涛. CRISPR/Cas9系统中的脱靶效应及检测技术研究进展[J]. 生物技术通报, 2020, 36(3): 78-87.

ZHANG Chen, LEI Zhan, LI Kai, SHANG Ying, XU Wen-tao. Research Progress on Off-target Effects and Detection Techniques in CRISPR/Cas9 Systems[J]. Biotechnology Bulletin, 2020, 36(3): 78-87.

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