Increasing the efficiency of CRISPR/Cas9-mediated precise genome editing in rats by inhibiting NHEJ and using Cas9 protein

doi:10.1080/15476286.2016.1185591

	Increasing the efficiency of CRISPR/Cas9-mediated precise genome editing in rats by inhibiting NHEJ and using Cas9 protein
	Ma, Yuanwu 1; Chen, Wei 1; Zhang, Xu 1; Yu, Lei 1; Dong, Wei 1; Pan, Shuo 1; Gao, Shan 1; Huang, Xingxu2 ; Zhang, Lianfeng 1
	2016
发表期刊	RNA BIOLOGY
ISSN	1547-6286
卷号	13 期号:7 页码:605-612
发表状态	已发表
DOI	10.1080/15476286.2016.1185591
摘要	Precise modifications such as site mutation, codon replacement, insertion or precise targeted deletion are needed for studies of accurate gene function. The CRISPR/Cas9 system has been proved as a powerful tool to generate gene knockout and knockin animals. But the homologous recombination (HR)-directed precise genetic modification mediated by CRISPR/Cas9 is relatively lower compared with nonhomologous end-joining (NHEJ) pathway and extremely expected to be improved. Here, in this study 2 strategies were used to increase the precise genetic modification in rats. Scr7, a DNA ligase IV inhibitor, first identified as an anti-cancer compound, and considered as a potential NHEJ inhibitor, was used to increase the HR-mediated precise genetic modification. Meanwhile, the Cas9 protein instead of mRNA was used to save the mRNA to protein translation step to improve the precise modification efficiency. The Fabp2 and Dbndd1 loci were selected to knockin Cre and CreERT2, respectively. Our result showed that both Scr7 and Cas9 protein can increase the precise modification.
关键词	CRISPR cas9 cre dbndd1 fabp2 homologous recombination (HR) rat
收录类别	SCI
语种	英语
资助项目	National Natural Science Foundation of China[31501001]
WOS研究方向	Biochemistry & Molecular Biology
WOS类目	Biochemistry & Molecular Biology
WOS记录号	WOS:000382959500001
出版者	TAYLOR & FRANCIS INC
WOS关键词	ZINC-FINGER NUCLEASES ; EMBRYO MICROINJECTION ; KNOCKOUT RATS ; MICE ; ENDONUCLEASE ; GENERATION ; REPAIR ; CELLS
原始文献类型	Article
引用统计
文献类型	期刊论文
条目标识符	https://kms.shanghaitech.edu.cn/handle/2MSLDSTB/2023
专题	生命科学与技术学院_PI研究组_黄行许组
通讯作者	Huang, Xingxu; Zhang, Lianfeng
作者单位	1.Chinese Acad Med Sci, Inst Lab Anim Sci, Key Lab Human Dis Comparat Med, Minist Hlth, Beijing, Peoples R China 2.ShanghaiTech Univ, Sch Life Sci & Technol, Shanghai, Peoples R China
通讯作者单位	生命科学与技术学院
推荐引用方式 GB/T 7714	Ma, Yuanwu,Chen, Wei,Zhang, Xu,et al. Increasing the efficiency of CRISPR/Cas9-mediated precise genome editing in rats by inhibiting NHEJ and using Cas9 protein[J]. RNA BIOLOGY,2016,13(7):605-612.
APA	Ma, Yuanwu.,Chen, Wei.,Zhang, Xu.,Yu, Lei.,Dong, Wei.,...&Zhang, Lianfeng.(2016).Increasing the efficiency of CRISPR/Cas9-mediated precise genome editing in rats by inhibiting NHEJ and using Cas9 protein.RNA BIOLOGY,13(7),605-612.
MLA	Ma, Yuanwu,et al."Increasing the efficiency of CRISPR/Cas9-mediated precise genome editing in rats by inhibiting NHEJ and using Cas9 protein".RNA BIOLOGY 13.7(2016):605-612.