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利用TALEN和CRISPR/Cas对水稻进行基因编辑

Gene Editing With TALEN and CRISPR/Cas in Rice.

作者信息

Bi Honghao, Yang Bing

机构信息

Iowa State University, Ames, IA, United States.

Iowa State University, Ames, IA, United States.

出版信息

Prog Mol Biol Transl Sci. 2017;149:81-98. doi: 10.1016/bs.pmbts.2017.04.006. Epub 2017 May 24.

Abstract

Engineered, site-specific nucleases induce genomic double-strand DNA breaks and break repair processes enable genome editing in a plethora of eukaryotic genomes. TALENs (transcription activator-like effector nucleases) and CRISPR/Cas (clustered regularly interspaced short palindromic repeats and CRISPR-associated proteins) are potent biotechnological tools used for genome editing. In rice, species-tailored editing tools have proven to be efficient and easy to use. Both tools are capable of generating DNA double-strand breaks (DSBs) in vivo and such breaks can be repaired either by error-prone NHEJ (nonhomologous end joining) that leads to nucleotide insertions or deletions or by HDR (homology-directed repair) if an appropriate exogenous DNA template is provided. NHEJ repair often results in gene knockout, while HDR results in precise nucleotide sequence or gene replacement. In this review, we revisit the molecular mechanisms underlying DSB repair in eukaryotes and review the TALEN and CRISPR technologies (CRISPR/Cas9, CRISPR/Cpf1, and Base Editor) developed and utilized for genome editing by scientists in rice community.

摘要

工程化的位点特异性核酸酶可诱导基因组双链DNA断裂,而断裂修复过程能够在众多真核生物基因组中实现基因组编辑。转录激活样效应因子核酸酶(TALENs)和规律成簇间隔短回文重复序列及其相关蛋白(CRISPR/Cas)是用于基因组编辑的强大生物技术工具。在水稻中,针对特定物种设计的编辑工具已被证明高效且易于使用。这两种工具都能够在体内产生DNA双链断裂(DSBs),并且如果提供合适的外源DNA模板,此类断裂可以通过易出错的非同源末端连接(NHEJ)进行修复,从而导致核苷酸插入或缺失,或者通过同源定向修复(HDR)进行修复。NHEJ修复通常会导致基因敲除,而HDR则会导致精确的核苷酸序列或基因替换。在本综述中,我们重新审视了真核生物中DSB修复的分子机制,并回顾了水稻研究领域的科学家们开发和应用于基因组编辑的TALEN和CRISPR技术(CRISPR/Cas9、CRISPR/Cpf1和碱基编辑器)。

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