单碱基编辑工具—腺嘌呤碱基编辑器ABE的研究进展

doi:10.3969/j.issn.1674-7968.2019.10.013

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摘要基因编辑技术CRISPR/Cas9自诞生以来,在基因敲除、敲入、碱基修复等领域得到了广泛的应用,但其效率低下,且存在非靶向切割,安全性较低,极大地限制了传统CRISPR/Cas9在高分辨率单碱基编辑中的应用。基于传统CRISPR/Cas9发展起来的碱基编辑器(base editor, BE)的出现则克服了这些弊端,其中,胞嘧啶碱基编辑器(cytosine base editor, CBE)能够使C•G转换为T•A,腺嘌呤碱基编辑器(adenine base editor, ABE)能够使A•T转换为G•C,都可以在不引入双链断裂的情况下实现高效的单碱基置换,这就避免了传统的CRISPR/Cas9在进行非同源末端连接(nonhomologous end-joining, NHEJ)时引入不可控的插入或缺失突变(Indels)。人类(Homo sapiens)大部分遗传疾病是由于碱基突变造成的,而单碱基编辑工具的出现能够在一定程度上纠正一定比例的致病性SNP,因此,其在动物模型构建、功能基因组学研究、分子育种、临床医学、转化医学等领域应用前景广阔。本文针对发展较晚、脱靶效率更低的腺嘌呤碱基编辑器ABE的原理、发展历程、应用和所面临的机遇与挑战进行了综述,以期为单碱基编辑技术的研究与应用提供参考。

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	李广栋
	张鲁
	富俊才
	连正兴
	刘国世

关键词 ：单碱基修复, 腺嘌呤碱基编辑器(ABE), 基因编辑

Abstract：Since the birth of the gene editing technology CRISPR/Cas9, it has been widely used in the fields of gene knockout, knock-in, and base repair. But its inefficiency and off-target cutting, low security, greatly limits the traditional CRISPR/Cas9 application in high-resolution single-base editing. The emergence of the base editor (BE) based on traditional CRISPR/Cas9 overcomes these drawbacks. The cytosine base editor (CBE) converts C•G to T•A, adenine base editor (ABE) enables the conversion of A•T to G•C, enabling efficient single base substitutions without introducing double-strand breaks, which avoids an uncontrollable insertion or deletion mutation (Indels) induced by the traditional CRISPR/Cas9 non-homologous end joining (NHEJ). Most of the genetic diseases in human are caused by base mutations, and the emergence of single-base editing tools can correct a certain proportion of pathogenic SNPs to some extent. Therefore it has broad application prospects in animal model construction, functional genomics research, molecular breeding, clinical medicine, and translational medicine. In this paper, the principle, development, application, opportunities and challenges of the just emerging ABEs with lower off-target efficiency are reviewed, in order to provide reference for the research and application of single-base editing technology.

Key words： Single base correction Adenine base editor (ABE) Gene editing

收稿日期: 2019-03-30

ZTFLH:

S-03

基金资助:国家转基因生物新品种培育科技重大专项(No. 2018ZX0800801B; No. 2016ZX08008003); 国家自然科学基金重点项目(No. 31830091); 北京市奶牛创新团队(BAIC06-2017)

通讯作者: gshliu@cau.edu.cn

引用本文:

李广栋, 张鲁, 富俊才, 连正兴, 刘国世. 单碱基编辑工具—腺嘌呤碱基编辑器ABE的研究进展[J]. 农业生物技术学报, 2019, 27(10): 1831-1839.
LI Guang-Dong, ZHANG Lu, FU Jun-Cai, LIAN Zheng-Xing, LIU Guo-Shi. Advances in Research on Single Base Editing Tool--Adenine Base Editor ABE. 农业生物技术学报, 2019, 27(10): 1831-1839.

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http://journal05.magtech.org.cn/Jwk_ny/CN/10.3969/j.issn.1674-7968.2019.10.013 或 http://journal05.magtech.org.cn/Jwk_ny/CN/Y2019/V27/I10/1831

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