Transcription-coupled donor DNA expression increases homologous recombination for efficient genome editing

doi:10.1093/nar/gkac676

	Transcription-coupled donor DNA expression increases homologous recombination for efficient genome editing
	Gao, Kaixuan; Zhang, Xuedi; Zhang, Zhenwu; Wu, Xiangyu; Guo, Yan; Fu, Pengchong; Sun, Angyang; Peng, Ju; Zheng, Jie; Yu, Pengfei; Wang, Tengfei; Ye, Qinying; Jiang, Jingwei; Wang, Haopeng; Lin, Chao-Po; Gao, Guanjun
	2022-08
发表期刊	NUCLEIC ACIDS RESEARCH (IF:16.6[JCR-2023],16.1[5-Year])
ISSN	0305-1048
EISSN	1362-4962
发表状态	已发表
DOI	10.1093/nar/gkac676
摘要	Genomes can be edited by homologous recombination stimulated by CRISPR/Cas9 [clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR-associated peptide 9]-induced DNA double-strand breaks. However, this approach is inefficient for inserting or deleting long fragments in mammalian cells. Here, we describe a simple genome-editing method, termed transcription-coupled Cas9-mediated editing (TEd), that can achieve higher efficiencies than canonical Cas9-mediated editing (CEd) in deleting genomic fragments, inserting/replacing large DNA fragments and introducing point mutations into mammalian cell lines. We also found that the transcription on DNA templates is crucial for the promotion of homology-directed repair, and that tethering transcripts from TEd donors to targeted sites further improves editing efficiency. The superior efficiency of TEd for the insertion and deletion of long DNA fragments expands the applications of CRISPR for editing mammalian genomes.
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收录类别	SCI ; SCIE
语种	英语
资助项目	National Natural Science Foundation of China["32070846","91740107"] ; National Postdoctoral Fellowship[2019M661652] ; Science, and Technology Commission of Shanghai Municipality[19JC1413600]
WOS研究方向	Biochemistry & Molecular Biology
WOS类目	Biochemistry & Molecular Biology
WOS记录号	WOS:000836131900001
出版者	OXFORD UNIV PRESS
引用统计	被引频次：7[WOS] [WOS记录] [WOS相关记录]
文献类型	期刊论文
条目标识符	https://kms.shanghaitech.edu.cn/handle/2MSLDSTB/214765
专题	生命科学与技术学院_硕士生生命科学与技术学院_PI研究组_林照博组生命科学与技术学院_PI研究组_王皞鹏组生命科学与技术学院_PI研究组_高冠军组生命科学与技术学院_博士生
通讯作者	Gao, Guanjun
作者单位	ShanghaiTech Univ, Gene Editing Ctr, Sch Life Sci & Technol, Shanghai 201210, Peoples R China
第一作者单位	生命科学与技术学院
通讯作者单位	生命科学与技术学院
第一作者的第一单位	生命科学与技术学院
推荐引用方式 GB/T 7714	Gao, Kaixuan,Zhang, Xuedi,Zhang, Zhenwu,et al. Transcription-coupled donor DNA expression increases homologous recombination for efficient genome editing[J]. NUCLEIC ACIDS RESEARCH,2022.
APA	Gao, Kaixuan.,Zhang, Xuedi.,Zhang, Zhenwu.,Wu, Xiangyu.,Guo, Yan.,...&Gao, Guanjun.(2022).Transcription-coupled donor DNA expression increases homologous recombination for efficient genome editing.NUCLEIC ACIDS RESEARCH.
MLA	Gao, Kaixuan,et al."Transcription-coupled donor DNA expression increases homologous recombination for efficient genome editing".NUCLEIC ACIDS RESEARCH (2022).