Spermatogenic Cell-Specific Gene Mutation in Mice via CRISPR-Cas9

查看全文 作　　者：Meizhu [1,2,3]Bai;Dan [2,3]Liang;Yinghua [2,3]Wang;Qing [2,3]Li;Yuxuan [2,3]Wu;Jinsong [1,2,3]Li

机构地区：[1]School of Life Science and Technology,Shanghai Tech University;[2]Group of Epigenetic Reprogramming,State Key Laboratory of Cell Biology,CAS Center for Excellence in Molecular Cell Science,Institute of Biochemistry and Cell Biology,Shanghai Institutes for Biological Sciences,Chinese Academy of Sciences;[3]Shanghai Key Laboratory of Molecular Andrology,Institute of Biochemistry and Cell Biology,Shanghai Institutes for Biological Sciences,Chinese Academy of Sciences

出　　处：《Journal of Genetics and Genomics》索引2016年第43卷第5期,共8页

基　　金：supported by the grants from the Ministry of Science and Technology of China(Nos.2014CB964803 and 2015AA020307);the National Natural Science Foundation of China(Nos.91319310,31225017 and 31530048);the Chinese Academy of Sciences(No.XDA01010403) 摘　　要：Tissue-specific knockout technology enables the analysis of the gene function in specific tissues in adult mammals.However,conventional strategy for producing tissue-specific knockout mice is a time- and labor-consuming process,restricting rapid study of the gene function in vivo.CRISPR-Cas9 system from bacteria is a simple and efficient gene-editing technique,which has enabled rapid generation of gene knockout lines in mouse by direct injection of CRISPR-Cas9 into zygotes.Here,we demonstrate CRISPR-Cas9-mediated spermatogenic cell-specific disruption of Scp3 gene in testes in one step.We first generated transgenic mice by pronuclear injection of a plasmid containing Hspa2 promoter driving Cas9 expression and showed Cas9 specific expression in spermatogenic cells.We then produced transgenic mice carrying Hspa2 promoter driven Cas9 and constitutive expressed sgRNA targeting Scp3 gene.Male founders were infertile due to developmental arrest of spermatogenic cells while female founders could produce progeny normally.Consistently,male progeny from female founders were infertile and females could transmit the transgenes to the next generation.Our study establishes a CRISPR-Cas9-based one-step strategy to analyze the gene function in adult tissues by a temporal-spatial pattern. 关键词：转基因小鼠基因突变细胞特异性基因功能分析组织特异性成年哺乳动物基因敲除编辑技术

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Spermatogenic Cell-Specific Gene Mutation in Mice via CRISPR-Cas9

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