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题名: Efficient ligase 3-dependent microhomology-mediated end joining repair of DNA double-strand breaks in zebrafish embryos
作者: He, Mu-Dan1, 2; Zhang, Feng-Hua1, 2; Wang, Hua-Lin1, 2; Wang, Hou-Peng1; Zhu, Zuo-Yan1; Sun, Yong-Hua1
关键词: Double-strand breaks repair ; Transcription activator-like effector nuclease (TALEN) ; CRISPR (clustered regularly interspaced short palindromic repeats)/Cas9 ; Microhomology-mediated end joining (MMEJ) ; Ligase 3 ; Ligase 4
刊名: MUTATION RESEARCH-FUNDAMENTAL AND MOLECULAR MECHANISMS OF MUTAGENESIS
发表日期: 2015-10-01
DOI: 10.1016/j.mrfmmm.2015.08.004
卷: 780, 页:86-96
收录类别: SCI
文章类型: Article
WOS标题词: Science & Technology ; Life Sciences & Biomedicine
类目[WOS]: Biotechnology & Applied Microbiology ; Genetics & Heredity ; Toxicology
研究领域[WOS]: Biotechnology & Applied Microbiology ; Genetics & Heredity ; Toxicology
英文摘要: DNA double-strand break (DSB) repair is of considerable importance for genomic integrity. Homologous recombination (HR) and non-homologous end joining (NHEJ) are considered as two major mechanistically distinct pathways involved in repairing DSBs. In recent years, another DSB repair pathway, namely, microhomology-mediated end joining (MMEJ), has received increasing attention. MMEJ is generally believed to utilize an alternative mechanism to repair DSBs when NHEJ and other mechanisms fail. In this study, we utilized zebrafish as an in vivo model to study DSB repair and demonstrated that efficient MMEJ repair occurred in the zebrafish genome when DSBs were induced using TALEN (transcription activator-like effector nuclease) or CRISPR (clustered regularly interspaced short palindromic repeats)/Cas9 technologies. The wide existence of MMEJ repair events in zebrafish embryos was further demonstrated via the injection of several in vitro-designed exogenous MMEJ reporters. Interestingly, the inhibition of endogenous ligase 4 activity significantly increased MMEJ frequency, and the inhibition of ligase 3 activity severely decreased MMEJ activity. These results suggest that MMEJ in zebrafish is dependent on ligase 3 but independent of ligase 4. This study will enhance our understanding of the mechanisms of MMEJ in vivo and facilitate inducing desirable mutations via DSB-induced repair. (C) 2015 Elsevier B.V. All rights reserved.
关键词[WOS]: GUIDED CAS9 NUCLEASE ; HOMOLOGOUS RECOMBINATION ; SACCHAROMYCES-CEREVISIAE ; VERTEBRATE CELLS ; KU80-DEFICIENT CELLS ; MAMMALIAN-CELLS ; GENE-EXPRESSION ; PATHWAY ; IV ; FISH
语种: 英语
WOS记录号: WOS:000362306000010
ISSN号: 0027-5107
Citation statistics:
内容类型: 期刊论文
URI标识: http://ir.ihb.ac.cn/handle/342005/27266
Appears in Collections:鱼类生物学及渔业生物技术研究中心_期刊论文

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作者单位: 1.Chinese Acad Sci, Inst Hydrobiol, State Key Lab Freshwater Ecol & Biotechnol, Wuhan 430072, Peoples R China
2.Univ Chinese Acad Sci, Beijing 100049, Peoples R China
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