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基因组编辑技术经历了第一代锌指核酸酶(zinc-finger nuclease, ZFNs)、第二代转录激活子样效应因子核酸酶(transcription activator-like effector nuclease, TALENs)的发展，以及当前应用最广泛、研究最深入的第三代规律性重复短回文序列簇(clustered regularly interspaced short palindromic repeat, CRISPR/Cas)系统。CRISPR/Cas系统可以在动物、植物、微生物基因组上实现定点敲除、单碱基替换和插入、小片段精准插入和缺失等多种功能。但目前最广泛使用的Cas9和Cas12a核酸酶的尺寸过大，以至于超过了很多载体的装载量而难以递送至细胞核。插入序列类似Cas9的B蛋白(insertion sequences Cas9-like B, IscB)和转座子相关转座蛋白B(transposon-associated transposase B, TnpB)等新型核酸酶，是IS200/IS605转座子超家族的成员，分别是Cas9和Cas12的祖先，两者的大小均约400个氨基酸，较小的体积使得它们在递送到动植物细胞核上有较大的优势，从而成为当前基因组编辑技术的研究热点之一。本文对IscB和TnpB的系统进化关系、作用原理、最新研究进展进行综述，以期为进行相关研究的人员提供帮助。
Abstract：Genome editing technology has evolved through the first generation of site-specific zinc finger nucleases(ZFNs), the second generation of transcription activator-like effector nucleases(TALENs), and currently the widely applied and extensively studied clustered regularly interspaced short palindromic repeat(CRISPR/Cas) systems. CRISPR/Cas system enables precise manipulation of genomes in animals, plants, and microorganisms, including site-specific deletions, single nucleotide substitutions, insertions, and precise incorporation of small fragments. However, CRISPR proteins such as Cas9 and Cas12a face challenges related to their large protein size, which hinders efficient translocation into the cell nucleus. Novel nucleases such as IscB and TnpB, members of the IS200/IS605 transposase superfamily, have emerged to address this issue. IscB serves as the ancestor of Cas9, while TnpB represents the ancestor of Cas12. Both have a structure of approximately 400 amino acids, and their reduced size provides a notable advantage in facilitating their delivery into the nuclei of both animal and plant cells, it has become one of the research hotspots of gene editing technology at present. In this paper, system evolutionary relationships, function principle and latest research progress of insertion sequences Cas9-like B(IscB) and transposon-associated transposase B(TnpB) are reviewed, in order to provide help for the relevant researchers.
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基本信息:
DOI：10.13417/j.gab.042.001267
中图分类号:Q78
引用信息:
[1]柴楠,刘雨馨,张瑞祥,等.基因组编辑工具的发展：从CRISPR/Cas9到TnpB[J].基因组学与应用生物学,2023,42(12):1267-1274.DOI:10.13417/j.gab.042.001267.
基金信息:
国家重点研发计划项目(2022YFF1002802); 国家自然科学基金面上项目(32272120)共同资助
2023-11-21
2023-11-21
2023-11-21