Volume 27, No 6, Jun 2017

ISSN: 1001-0602 
EISSN: 1748-7838 2018 
impact factor 17.848* 
(Clarivate Analytics, 2019)

Volume 27 Issue 6, June 2017: 815-829

ORIGINAL ARTICLES

CRISPR-Cas9-mediated genome editing in one blastomere of two-cell embryos reveals a novel Tet3 function in regulating neocortical development

Lingbo Wang^1,*, Min-Yin Li^2,*, Chao Qu¹, Wan-Ying Miao², Qi Yin¹, Jiaoyang Liao¹, Hua-Teng Cao², Min Huang^3,4, Kai Wang¹, Erwei Zuo¹, Guangdun Peng⁵, Shu-Xin Zhang², Guodong Chen⁶, Qing Li¹, Ke Tang⁷, Qian Yu8, Zhoujie Li⁸, Catherine CL Wong^3,4, Guoliang Xu⁶, Naihe Jing⁵, Xiang Yu² and Jinsong Li¹

¹State Key Laboratory of Cell Biology, Shanghai Key Laboratory of Molecular Andrology, CAS Center for Excellence in Molecular Cell Science, Institute of Biochemistry and Cell Biology, Chinese Academy of Sciences, University of Chinese Academy of Sciences, 320 Yueyang Road, Shanghai 200031, China
²Institute of Neuroscience and State Key Laboratory of Neuroscience, CAS Center for Excellence in Brain Science and Intelligence Technology, Chinese Academy of Sciences, University of Chinese Academy of Sciences, 320 Yueyang Road, Shanghai 200031, China
³National Center for Protein Science Shanghai, Institute of Biochemistry and Cell Biology, Chinese Academy of Sciences, 333 Haike Road, Shanghai 201203, China
⁴Shanghai Science Research Center, Chinese Academy of Sciences, Shanghai 201204, China
⁵State Key Laboratory of Cell Biology, Institute of Biochemistry and Cell Biology, Chinese Academy of Sciences, University of Chinese Academy of Sciences, 320 Yueyang Road, Shanghai 200031, China
⁶State Key Laboratory of Molecular Biology, Shanghai Key Laboratory of Molecular Andrology, Institute of Biochemistry and Cell Biology, Chinese Academy of Sciences, University of Chinese Academy of Sciences, 320 Yueyang Road, Shanghai 200031, China
⁷Institute of Life Science, Nanchang University, Nanchang, Jiangxi 330031, China
⁸Animal Core Facility, Institute of Biochemistry and Cell Biology, Chinese Academy of Sciences, Shanghai 200031, China Correspondence: Jinsong Li, E-mail: jsli@sibcb.ac.cn; Xiang Yu,(yuxiang@ion.ac.cn)

Studying the early function of essential genes is an important and challenging problem in developmental biology. Here, we established a method for rapidly inducing CRISPR-Cas9-mediated mutations in one blastomere of two-cell stage embryos, termed 2-cell embryo-CRISPR-Cas9 injection (2CC), to study the in vivo function of essential (or unknown) genes in founder chimeric mice. By injecting both Cre mRNA and CRISPR-Cas9 targeting the gene of interest into fluorescent reporter mice, the 2CC method can trace both wild-type and mutant cells at different developmental stages, offering internal control for phenotypic analyses of mutant cells. Using this method, we identified novel functions of the essential gene Tet3 in regulating excitatory and inhibitory synaptic transmission in the developing mouse cerebral cortex. By generating chimeric mutant mice, the 2CC method allows for the rapid screening of gene function in multiple tissues and cell types in founder chimeric mice, significantly expanding the current armamentarium of genetic tools.

10.1038/cr.2017.58

FULL TEXT | PDF

Browse 1950