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Efficient Generation of Gene-Modified Pigs Harboring Precise Orthologous Human Mutation via CRISPR/Cas9-Induced Homology-Directed Repair in Zygotes | |
2016-01 | |
Source Publication | HUMAN MUTATION
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ISSN | 1059-7794 |
Volume | 37Issue:1Pages:110-118 |
Status | 已发表 |
DOI | 10.1002/humu.22913 |
Abstract | Precise genetic mutation of model animals is highly valuable for functional investigation of human mutations. Clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR-associated 9 (Cas9)-induced homology-directed repair (HDR) is usually used for precise genetic mutation, being limited by the relatively low efficiency compared with that of non-homologous end joining (NHEJ). Although inhibition of NHEJ was shown to enhance HDR-derived mutation, in this work, without inhibition of NHEJ, we first generated gene-modified pigs harboring precise orthologous human mutation (Sox10 c.A325>T) via CRISPR/Cas9-induced HDR in zygotes using single-strand oligo DNA (ssODN) as template with an efficiency as high as 80%, indicating that pig zygotes exhibited high activities of HDR relative to NHEJ and were highly amendable to genetic mutation via CIRSPR/Cas9-induced HDR. Besides, we found a higher concentration of ssODN remarkably reduced HDR-derived mutation in pig zygotes, suggesting a possible balance for optimal HDR-derived mutation in zygotes between the excessive accessibility to HDR templates and the activities of HDR relative to NHEJ which appeared to be negatively correlated to ssODN concentration. In addition, the HDR-derived mutation, as well as those from NHEJ, extensively integrated into various tissues including gonad of founder pig without detected off-targeting, suggesting CRISPR/Cas9-induced HDR in zygotes is a reliable approach for precise genetic mutation in pigs. |
Keyword | SOX10 CRISPR Cas9 homology-directed repair pigs model organism disease model |
Indexed By | SCI |
Language | 英语 |
Funding Project | National Major Scientific Projects Specifically for R&D of Transgenic Organisms[2014ZX08009-003-006] |
WOS Research Area | Genetics & Heredity |
WOS Subject | Genetics & Heredity |
WOS ID | WOS:000369159900011 |
Publisher | WILEY |
WOS Keyword | CONDITIONAL ALLELES ; CLONING EFFICIENCY ; KNOCKOUT RATS ; NEURAL CREST ; SOX10 ; SPECIFICITY ; CLEAVAGE |
Original Document Type | Article |
Citation statistics | |
Document Type | 期刊论文 |
Identifier | https://kms.shanghaitech.edu.cn/handle/2MSLDSTB/2053 |
Collection | 生命科学与技术学院_PI研究组_黄行许组 |
Corresponding Author | Wang, Yong; Wei, Hong |
Affiliation | 1.Third Mil Med Univ, Coll Basic Med Sci, Dept Lab Anim Sci, Chongqing, Peoples R China 2.ShanghaiTech Univ, Sch Life Sci & Technol, Shanghai, Peoples R China |
Recommended Citation GB/T 7714 | Zhou, Xiaoyang,Wang, Lulu,Du, Yinan,et al. Efficient Generation of Gene-Modified Pigs Harboring Precise Orthologous Human Mutation via CRISPR/Cas9-Induced Homology-Directed Repair in Zygotes[J]. HUMAN MUTATION,2016,37(1):110-118. |
APA | Zhou, Xiaoyang.,Wang, Lulu.,Du, Yinan.,Xie, Fei.,Li, Liang.,...&Wei, Hong.(2016).Efficient Generation of Gene-Modified Pigs Harboring Precise Orthologous Human Mutation via CRISPR/Cas9-Induced Homology-Directed Repair in Zygotes.HUMAN MUTATION,37(1),110-118. |
MLA | Zhou, Xiaoyang,et al."Efficient Generation of Gene-Modified Pigs Harboring Precise Orthologous Human Mutation via CRISPR/Cas9-Induced Homology-Directed Repair in Zygotes".HUMAN MUTATION 37.1(2016):110-118. |
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