CRISPR/Cas9-based genome engineering of zebrafish using a seamless integration strategy | |
Luo, Juan-Juan1,2; Bian, Wan-Ping2; Liu, Yi2,3; Huang, Hai-Yang2; Yin, Qian2; Yang, Xiao-Jun1; Pei, De-Sheng2 | |
刊名 | FASEB JOURNAL |
2018-09-01 | |
卷号 | 32期号:9页码:5132-5142 |
关键词 | Gene Editing Mmej Transgenesis Knock-in |
ISSN号 | 0892-6638 |
DOI | 10.1096/fj.201800077RR |
英文摘要 | Numerous feasible methods for inserting large fragments of exogenous DNA sequences into the zebrafish genome have been developed, as has genome editing technology using programmable nucleases. However, the coding sequences of targeted endogenous genes are disrupted, and the expression patterns of inserted exogenous genes cannot completely recapitulate those of endogenous genes. Here we describe the establishment of a novel strategy for endogenous promoter-driven and microhomology-mediated end-joining-dependent integration of a donor vector using clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR-associated (Cas) 9. We successfully integrated mCherry into the final coding sequence of targeted genes to generate seamless transgenic zebrafish lines with high efficiency. This novel seamless transgenesis technique not only maintained the integrity of the endogenous gene but also did not disrupt the function of targeted gene. Therefore, our microhomology-mediated end-joining-mediated transgenesis strategy may have broader applications in gene therapy. Moreover, this novel seamless gene-editing strategy in zebrafish provides a valuable new transgenesis technique, which was driven by endogenous promoters and in vivo animal reporter modes for translational medicine. It is expected to be a standard gene-editing technique in the field of zebrafish, leading to some important breakthroughs for studies in early embryogenesis.Luo, J.-J., Bian, W.-P., Liu, Y., Huang, H.-Y., Yin, Q., Yang, X.-J., Pei, D.-S. CRISPR/Cas9-based genome engineering of zebrafish using a seamless integration strategy. |
资助项目 | Three Hundred Leading Talents in Scientific and Technological Innovation Program of Chongqing[CSTCCXLJRC201714] ; National Natural Science Foundation of China[81572494] ; Key Application and Development Program of the Chongqing Science and Technology Commission[cstc2014yykfC20004] ; Key Application and Development Program of the Chongqing Science and Technology Commission[cstc2014yykfC20002] ; Natural Science Foundation of Guangdong Province[2015A030313440] ; Hundred Talents Program of the Chinese Academy of Sciences |
WOS研究方向 | Biochemistry & Molecular Biology ; Life Sciences & Biomedicine - Other Topics ; Cell Biology |
语种 | 英语 |
出版者 | FEDERATION AMER SOC EXP BIOL |
WOS记录号 | WOS:000443817700041 |
内容类型 | 期刊论文 |
源URL | [http://119.78.100.138/handle/2HOD01W0/6663] |
专题 | 中国科学院重庆绿色智能技术研究院 |
作者单位 | 1.Shantou Univ, Ctr Neurosci, Coll Med, 22 Xinling Rd, Shantou 515041, Peoples R China 2.Chinese Acad Sci, Chongqing Inst Green & Intelligent Technol, Chongqing 400714, Peoples R China 3.Univ Chinese Acad Sci, Beijing, Peoples R China |
推荐引用方式 GB/T 7714 | Luo, Juan-Juan,Bian, Wan-Ping,Liu, Yi,et al. CRISPR/Cas9-based genome engineering of zebrafish using a seamless integration strategy[J]. FASEB JOURNAL,2018,32(9):5132-5142. |
APA | Luo, Juan-Juan.,Bian, Wan-Ping.,Liu, Yi.,Huang, Hai-Yang.,Yin, Qian.,...&Pei, De-Sheng.(2018).CRISPR/Cas9-based genome engineering of zebrafish using a seamless integration strategy.FASEB JOURNAL,32(9),5132-5142. |
MLA | Luo, Juan-Juan,et al."CRISPR/Cas9-based genome engineering of zebrafish using a seamless integration strategy".FASEB JOURNAL 32.9(2018):5132-5142. |
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