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Efficient generation of zebrafish maternal-zygotic mutants through transplantation of ectopically induced and Cas9/gRNA targeted primordial germ cells
Zhang, Fenghua1,2; Li, Xianmei1,2; He, Mudan1,2; Ye, Ding1,2; Xiong, Feng1,2; Amin, Golpour1,2; Zhu, Zuoyan1,2; Sun, Yonghua1,2
2020-01-20
Source PublicationJOURNAL OF GENETICS AND GENOMICS
ISSN1673-8527
Volume47Issue:1Pages:37-47
Abstract

The clustered regularly interspaced short palindromic repeats (CRISPR)/Cas9 technology has been widely utilized for knocking out genes involved in various biological processes in zebrafish. Despite this technology is efficient for generating different mutations, one of the main drawbacks is low survival rate during embryogenesis when knocking out some embryonic lethal genes. To overcome this problem, we developed a novel strategy using a combination of CRISPR/Cas9 mediated gene knockout with primordial germ cell (PGC) transplantation (PGCT) to facilitate and speed up the process of zebrafish mutant generation, particularly for embryonic lethal genes. Firstly, we optimized the procedure for CRISPR/Cas9 targeted PGCT by increasing the efficiencies of genome mutation in PGCs and induction of PGC fates in donor embryos for PGCT. Secondly, the optimized CRISPR/Cas9 targeted PGCT was utilized for generation of maternal-zygotic (MZ) mutants of tcf7l1a (gene essential for head development), pou5f3 (gene essential for zygotic genome activation) and chd (gene essential for dorsal development) at F-1 generation with relatively high efficiency. Finally, we revealed some novel phenotypes in MZ mutants of tcf7l1a and chd, as MZtcf7l1a showed elevated neural crest development while MZchd had much severer ventralization than its zygotic counterparts. Therefore, this study presents an efficient and powerful method for generating MZ mutants of embryonic lethal genes in zebrafish. It is also feasible to speed up the genome editing in commercial fishes by utilizing a similar approach by surrogate production of CRISPR/Cas9 targeted germ cells. Copyright (C) 2020, Institute of Genetics and Developmental Biology, Chinese Academy of Sciences, and Genetics Society of China. Published by Elsevier Limited and Science Press. All rights reserved.

KeywordZebrafish CRISPR/Cas9 Primordial germ cells Transplantation Maternal zygotic mutant
DOI10.1016/j.jgg.2019.12.004
Indexed BySCI
Language英语
WOS Research AreaBiochemistry & Molecular Biology ; Genetics & Heredity
WOS SubjectBiochemistry & Molecular Biology ; Genetics & Heredity
WOS IDWOS:000524095600004
WOS KeywordGENE-EXPRESSION ; MIDBLASTULA TRANSITION ; END
PublisherSCIENCE PRESS
Citation statistics
Cited Times:2[WOS]   [WOS Record]     [Related Records in WOS]
Document Type期刊论文
Identifierhttp://ir.ihb.ac.cn/handle/342005/36112
Collection鱼类生物学及渔业生物技术研究中心_期刊论文
Corresponding AuthorSun, Yonghua
Affiliation1.Chinese Acad Sci, Innovat Acad Seed Design, Inst Hydrobiol, State Key Lab Freshwater Ecol & Biotechnol, Wuhan 430072, Peoples R China
2.Univ Chinese Acad Sci, Coll Adv Agr Sci, Beijing 100049, Peoples R China
Recommended Citation
GB/T 7714
Zhang, Fenghua,Li, Xianmei,He, Mudan,et al. Efficient generation of zebrafish maternal-zygotic mutants through transplantation of ectopically induced and Cas9/gRNA targeted primordial germ cells[J]. JOURNAL OF GENETICS AND GENOMICS,2020,47(1):37-47.
APA Zhang, Fenghua.,Li, Xianmei.,He, Mudan.,Ye, Ding.,Xiong, Feng.,...&Sun, Yonghua.(2020).Efficient generation of zebrafish maternal-zygotic mutants through transplantation of ectopically induced and Cas9/gRNA targeted primordial germ cells.JOURNAL OF GENETICS AND GENOMICS,47(1),37-47.
MLA Zhang, Fenghua,et al."Efficient generation of zebrafish maternal-zygotic mutants through transplantation of ectopically induced and Cas9/gRNA targeted primordial germ cells".JOURNAL OF GENETICS AND GENOMICS 47.1(2020):37-47.
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