Creating a monomeric endonuclease TALE-I-SceI with high specificity and   low genotoxicity in human cells

doi:10.1093/nar/gku1339

CORC > 北京大学 > 生命科学学院

	Creating a monomeric endonuclease TALE-I-SceI with high specificity and low genotoxicity in human cells
	Lin, Jianfei ; Chen, He ; Luo, Ling ; Lai, Yongrong ; Xie, Wei ; Kee, Kehkooi
刊名	核酸研究
	2015
关键词	ZINC-FINGER NUCLEASES HISTONE H2AX PHOSPHORYLATION GENE-THERAPY MAMMALIAN-CELLS CAS9 ARCHITECTURE SYSTEMS REPAIR BREAKS
DOI	10.1093/nar/gku1339
英文摘要	To correct a DNA mutation in the human genome for gene therapy, homology-directed repair (HDR) needs to be specific and have the lowest off-target effects to protect the human genome from deleterious mutations. Zinc finger nucleases, transcription activatorlike effector nuclease (TALEN) and CRISPR-CAS9 systems have been engineered and used extensively to recognize and modify specific DNA sequences. Although TALEN and CRISPR/CAS9 could induce high levels of HDR in human cells, their genotoxicity was significantly higher. Here, we report the creation of a monomeric endonuclease that can recognize at least 33 bp by fusing the DNA-recognizing domain of TALEN (TALE) to a re-engineered homing endonuclease I-SceI. After sequentially re-engineering I-SceI to recognize 18 bp of the human beta-globin sequence, the re-engineered I-SceI induced HDR in human cells. When the re-engineered I-SceI was fused to TALE (TALE-ISVB2), the chimeric endonuclease induced the same HDR rate at the human beta-globin gene locus as that induced by TALEN, but significantly reduced genotoxicity. We further demonstrated that TALE-ISVB2 specifically targeted at the beta-globin sequence in human hematopoietic stem cells. Therefore, this monomeric endonuclease has the potential to be used in therapeutic gene targeting in human cells.; http://gateway.webofknowledge.com/gateway/Gateway.cgi?GWVersion=2&SrcApp=PARTNER_APP&SrcAuth=LinksAMR&KeyUT=WOS:000350209000042&DestLinkType=FullRecord&DestApp=ALL_WOS&UsrCustomerID=8e1609b174ce4e31116a60747a720701 ; Biochemistry & Molecular Biology; SCI(E); PubMed; 3; ARTICLE; kkee@tsinghua.edu.cn; 2; 1112-1122; 43
语种	英语
内容类型	期刊论文
源URL	[http://ir.pku.edu.cn/handle/20.500.11897/319917]
专题	生命科学学院
推荐引用方式 GB/T 7714	Lin, Jianfei,Chen, He,Luo, Ling,et al. Creating a monomeric endonuclease TALE-I-SceI with high specificity and low genotoxicity in human cells[J]. 核酸研究,2015.
APA	Lin, Jianfei,Chen, He,Luo, Ling,Lai, Yongrong,Xie, Wei,&Kee, Kehkooi.(2015).Creating a monomeric endonuclease TALE-I-SceI with high specificity and low genotoxicity in human cells.核酸研究.
MLA	Lin, Jianfei,et al."Creating a monomeric endonuclease TALE-I-SceI with high specificity and low genotoxicity in human cells".核酸研究 (2015).