基于CRISPR/Cas系统的单碱基编辑技术研究进展<sup>*</sup>

doi:10.13523/j.cb.2007051

中国生物工程杂志

2020, Vol. 40

Issue (12): 58-66 DOI: 10.13523/j.cb.2007051

综述

基于CRISPR/Cas系统的单碱基编辑技术研究进展^*

王玥,牟彦双(

),刘忠华

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

Progress of CRISPR/Cas Base Editing System

WANG Yue,MU Yan-shuang(

),LIU Zhong-hua

Heilongjiang Key Laboratory of Animal Cell and Genetic Engineering, Northeast Agricultural University, Harbin 150030, China

全文: PDF(30451 KB) HTML

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。

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

Abstract:

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.

Key words: CRISPR/Cas9 Gene editing Single base editing Cytosine Deaminase Adenosine deaminase

收稿日期: 2020-07-30 出版日期: 2021-01-14

ZTFLH:

Q789

基金资助: *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资助项目(LH2020C016)

通讯作者: 牟彦双 E-mail: muyanshuang@neau.edu.cn

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引用本文:

王玥,牟彦双,刘忠华. 基于CRISPR/Cas系统的单碱基编辑技术研究进展^*[J]. 中国生物工程杂志, 2020, 40(12): 58-66.

WANG Yue,MU Yan-shuang,LIU Zhong-hua. Progress of CRISPR/Cas Base Editing System. China Biotechnology, 2020, 40(12): 58-66.

链接本文:

https://manu60.magtech.com.cn/biotech/CN/10.13523/j.cb.2007051 或 https://manu60.magtech.com.cn/biotech/CN/Y2020/V40/I12/58

图1 CBE系统原理示意图

图2 ABE系统原理示意图

图3 双碱基编辑原理示意图

图4 RESCUE系统用pre-crRNA引导阵列对C到U和A到I进行多重编辑的示意图

图5 BE碱基编辑系统版本汇总

图6 CRISPR-X工作原理图

图7 dCpf1-BEs工作原理图

图8 eBE-S3结构示意图

图9 YE1-BE3-FNLS工作原理图

图10 hyCBE碱基编辑系统原理图

表1 ABE碱基编辑系统优化表

图11 ABE碱基编辑系统优化图

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