基于CRISPR/Cas系统的单碱基编辑技术研究进展  

Progress of CRISPR/Cas Base Editing System

作　　者：王玥[1] 牟彦双[1] 刘忠华[1] WANG Yue;MU Yan-shuang;LIU Zhong-hua(Heilongjiang Key Laboratory of Animal Cell and Genetie Engineering,Norheapt Agricultural University,Harbin 150030,China)

机构地区：[1]东北农业大学黑龙江省动物细胞与遗传工程重点实验室,哈尔滨150030

出　　处：《中国生物工程杂志》2020年第12期58-66,共9页China Biotechnology

基　　金：转基因生物新品种培育科技重大专项(2016ZX08009-003-006);国家重点研发计划(2016YFA0100200);黑龙江省自然科学基金联合引导项目(LH2020C016)资助项目。

摘　　要：基于CRISPR/Cas系统出现的单碱基编辑技术可以实现高效且简便的单个碱基的替换编辑,其原理是将胞嘧啶脱氨酶(cytosine deaminase)或腺苷脱氨酶(adenosine deaminase)与Cas9n(D10A)形成融合蛋白,通过CRISPR/Cas精准识别和定位DNA上的靶位点后,利用胞嘧啶脱氨酶或腺苷脱氨酶将靶点距离sgRNA位点基序(protospacer adjacent motif,PAM)序列端的4~7位的单个碱基发生单碱基转换或颠换。对基于CRISPR/Cas系统的单碱基编辑技术发现的历史、组成和分类、工作原理进行了概述,并总结了该系统最新进展及应用。Base editing is a precise genome editing technique based on the CRISPR/Cas system.Base editing use cytosine deaminases or adenine deaminases to perform base editing,and achieve base substitutions,respectively.Base editor,engineered by fusing the Cas9 nikase( Cas9n) with a cytidine deaminase enzyme is guided by a sgRNA to the target site.The position is the single base 4-7 of the target distance from the end of the sgRNA site motif( protospacer adjacent motif,PAM) sequence for editing.Without cutting doublestrand DNAs,base editor can precisely mediate the direct conversion of cytidine to thymineor or guanine to adenine.Therefore the base editing history,composition,working principle,and technology developments were briefly described.

关 键 词：CRISPR/Cas9 基因编辑 单碱基编辑 胞嘧啶脱氨酶 腺苷脱氨酶 

分 类 号：Q789[生物学—分子生物学]