单位：[1]Department of Neurobiology, Beijing Key Laboratory of Neural Regeneration and Repair, School of Basic Medical Sciences, Capital Medical University, Beijing100069, China[2]Molecular Laboratory for Gene Therapy and Tooth Regeneration, Beijing Key Laboratory of Tooth Regeneration and FunctionReconstruction, School of Stomatology, Capital Medical University, Beijing 100050, China[3]Laboratory Animal Center, Capital Medical University, Beijing100069, China[4]Experimental and Translational Research Center, Beijing Friendship Hospital, Capital Medical University, Beijing 100050, China医技科室北京市临床医学研究所实验中心首都医科大学附属北京友谊医院[5]Laboratory Animal Center, Peking University, Beijing 100871, China[6]Department of Oral Pathology, Beijing Stomatology Hospital, Capital MedicalUniversity, Beijing 100050, China[7]Department of Biochemistry and Molecular Biology, School of Basic Medical Sciences, Capital Medical University, Beijing100069, China
A substantial number of mouse genes, about 25%, are embryonically lethal when knocked out. Using current genetic tools, such as the CRISPR-Cas9 system, it is difficult-or even impossible-to produce viable mice with heritable embryonically lethal mutations. Here, we establish a one-step method for microinjection of CRISPR reagents into one blastomere of two-cell embryos to generate viable chimeric founder mice with a heritable embryonically lethal mutation, of either Virma or Dpm1. By examining founder mice, we identify a phenotype and role of Virma in regulating kidney metabolism in adult mice. Additionally, we generate knockout mice with a heritable postnatally lethal mutation, of either Slc17a5 or Ctla-4, and study its function in vivo. This one-step method provides a convenient system that rapidly generates knockout mice possessing lethal phenotypes. This allows relatively easy in vivo study of the associated genes' functions.
基金：
National Natural Science Foundation of ChinaNational Natural Science Foundation of China (NSFC) [91649124]; Beijing Municipality Government grants [PXM2018_014226_000021, PXM2017_014226_000023, PXM2018_193312_000006_0028S643_FCG PXM2016_014226_000034, PXM2016_014226_000034, PXM2016_014226_000006, PXM2015_014226_000116]
第一作者单位：[1]Department of Neurobiology, Beijing Key Laboratory of Neural Regeneration and Repair, School of Basic Medical Sciences, Capital Medical University, Beijing100069, China[2]Molecular Laboratory for Gene Therapy and Tooth Regeneration, Beijing Key Laboratory of Tooth Regeneration and FunctionReconstruction, School of Stomatology, Capital Medical University, Beijing 100050, China[3]Laboratory Animal Center, Capital Medical University, Beijing100069, China
共同第一作者：
通讯作者：
通讯机构：[1]Department of Neurobiology, Beijing Key Laboratory of Neural Regeneration and Repair, School of Basic Medical Sciences, Capital Medical University, Beijing100069, China[2]Molecular Laboratory for Gene Therapy and Tooth Regeneration, Beijing Key Laboratory of Tooth Regeneration and FunctionReconstruction, School of Stomatology, Capital Medical University, Beijing 100050, China[3]Laboratory Animal Center, Capital Medical University, Beijing100069, China[7]Department of Biochemistry and Molecular Biology, School of Basic Medical Sciences, Capital Medical University, Beijing100069, China
推荐引用方式(GB/T 7714)：
Yi Wu,Jing Zhang,Boya Peng,et al.Generating viable mice with heritable embryonically lethal mutations using the CRISPR-Cas9 system in two-cell embryos[J].NATURE COMMUNICATIONS.2019,10:doi:10.1038/s41467-019-10748-2.
APA：
Yi Wu,Jing Zhang,Boya Peng,Dan Tian,Dong Zhang...&Songlin Wang.(2019).Generating viable mice with heritable embryonically lethal mutations using the CRISPR-Cas9 system in two-cell embryos.NATURE COMMUNICATIONS,10,
MLA：
Yi Wu,et al."Generating viable mice with heritable embryonically lethal mutations using the CRISPR-Cas9 system in two-cell embryos".NATURE COMMUNICATIONS 10.(2019)
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