CRISPR-Cas9-mediated base-editing screening in mice identifies DND1 amino acids that are critical for primordial germ cell development

doi:10.1038/s41556-018-0202-4

	CRISPR-Cas9-mediated base-editing screening in mice identifies DND1 amino acids that are critical for primordial germ cell development
	Li, Qing 1; Li, Yanjing 2; Yang, Suming 1; Huang, Shuo3 ; Yan, Meng 1; Ding, Yifu 1; Tang, Wei 4; Lou, Xiwen 2; Yin, Qi 1; Sun, Zhanfei3 ; Lu, Lei 1,5; Shi, Huijuan 6; Wang, Hongyan 5; Chen, Yong2 ; Li, Jinsong1,3
	2018-11
发表期刊	NATURE CELL BIOLOGY (IF:17.3[JCR-2023],24.2[5-Year])
ISSN	1465-7392
卷号	20 期号:11 页码:1315-+
发表状态	已发表
DOI	10.1038/s41556-018-0202-4
摘要	CRISPR-mediated base editing can introduce single-nucleotide changes in the DNA of living cells. One intriguing application of base editing is to screen pivotal amino acids for protein function in vivo; however, it has not been achieved. Here, we report an enhanced third-generation base-editing system with extra nuclear localization sequences that can efficiently introduce a homozygous base mutation in embryonic stem cells. Meanwhile, we establish a strategy to generate base-mutant mice by injection of haploid embryonic stem cells carrying a constitutively expressed enhanced third-generation base-editing system (4B2N1) and single guide RNA into oocytes. Moreover, transfection of 4B2N1 cells with a single guide RNA library targeting the Dnd1 gene allows one-step generation of mutant mice with a base mutation. This enables the identification of four missense mutations that completely deplete primordial germ cells through disruption of DND1 protein stability and protein-protein interaction. Thus, our strategy provides an effective tool for in vivo screening of amino acids that are crucial for protein function.
收录类别	SCI ; SCIE
语种	英语
资助项目	Shanghai Municipal Commission for Science and Technology[16JC1420500] ; Shanghai Municipal Commission for Science and Technology[17JC1400900] ; Shanghai Municipal Commission for Science and Technology[17JC1420102] ; Shanghai Municipal Commission for Science and Technology[17411954900]
WOS研究方向	Cell Biology
WOS类目	Cell Biology
WOS记录号	WOS:000448320700013
出版者	NATURE PUBLISHING GROUP
WOS关键词	TER MUTATION ; GENOMIC DNA ; STEM-CELLS ; GENE ; GENERATION ; SITES ; COMPLEX
原始文献类型	Article
引用统计	正在获取...
文献类型	期刊论文
条目标识符	https://kms.shanghaitech.edu.cn/handle/2MSLDSTB/28237
专题	生命科学与技术学院_硕士生生命科学与技术学院_特聘教授组_陈勇组生命科学与技术学院_特聘教授组_李劲松组
通讯作者	Chen, Yong; Li, Jinsong
作者单位	1.Univ Chinese Acad Sci, Chinese Acad Sci, Shanghai Inst Biochem & Cell Biol,CAS Ctr Excelle, State Key Lab Cell Biol,Shanghai Key Lab Mol Andr, Shanghai, Peoples R China 2.Univ Chinese Acad Sci, Chinese Acad Sci,Shanghai Sci Res Ctr, Shanghai Inst Biochem & Cell Biol,Natl Ctr Prot S, CAS Ctr Excellence Mol Cell Sci,State Key Lab Mol, Shanghai, Peoples R China 3.Shanghai Tech Univ, Sch Life Sci & Technol, Shanghai, Peoples R China 4.Univ Chinese Acad Sci, Shanghai Inst Biochem & Cell Biol, Chinese Acad Sci, Anim Core Facil, Shanghai, Peoples R China 5.Fudan Univ, Inst Reprod & Dev, Hosp & Inst Obstet & Gynecol,Sch Life Sci, State Key Lab Genet Engn,Collaborat Innovat Ctr G, Shanghai, Peoples R China 6.Shanghai Inst Planned Parenthood Res, Natl Populat & Family Planning Comm, Key Lab Contracept Drugs & Devices, Shanghai, Peoples R China
通讯作者单位	生命科学与技术学院
推荐引用方式 GB/T 7714	Li, Qing,Li, Yanjing,Yang, Suming,et al. CRISPR-Cas9-mediated base-editing screening in mice identifies DND1 amino acids that are critical for primordial germ cell development[J]. NATURE CELL BIOLOGY,2018,20(11):1315-+.
APA	Li, Qing.,Li, Yanjing.,Yang, Suming.,Huang, Shuo.,Yan, Meng.,...&Li, Jinsong.(2018).CRISPR-Cas9-mediated base-editing screening in mice identifies DND1 amino acids that are critical for primordial germ cell development.NATURE CELL BIOLOGY,20(11),1315-+.
MLA	Li, Qing,et al."CRISPR-Cas9-mediated base-editing screening in mice identifies DND1 amino acids that are critical for primordial germ cell development".NATURE CELL BIOLOGY 20.11(2018):1315-+.