CRISPR-Cas9-mediated base-editing screening in mice identifies DND1 amino acids that are critical for primordial germ cell development | |
Li, Qing1; Yang, Suming1; Yan, Meng1; Ding, Yifu1; Yin, Qi1; Lu, Lei1,5; Li, Jinsong1,3; Li, Yanjing2; Lou, Xiwen2; Chen, Yong2 | |
刊名 | NATURE CELL BIOLOGY |
2018 | |
卷号 | 20期号:11页码:1315-+ |
关键词 | Embryonic Stem-cells Mouse Embryos Ter Mutation Dna Cleavage Genomic Dna Gene Generation Sites Complex Rnas |
ISSN号 | 1465-7392 |
DOI | 10.1038/s41556-018-0202-4 |
文献子类 | Article |
英文摘要 | 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. |
电子版国际标准刊号 | 1476-4679 |
WOS研究方向 | Cell Biology |
语种 | 英语 |
WOS记录号 | WOS:000448320700013 |
内容类型 | 期刊论文 |
版本 | 出版稿 |
源URL | [http://202.127.25.143/handle/331003/3387] |
专题 | 生化所2018年发文 上海生化细胞研究所_上海生科院生化细胞研究所 |
通讯作者 | Li, Jinsong; Chen, Yong |
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 |
推荐引用方式 GB/T 7714 | Li, Qing,Yang, Suming,Yan, Meng,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.,Yang, Suming.,Yan, Meng.,Ding, Yifu.,Yin, Qi.,...&Shi, Huijuan.(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-+. |
个性服务 |
查看访问统计 |
相关权益政策 |
暂无数据 |
收藏/分享 |
除非特别说明,本系统中所有内容都受版权保护,并保留所有权利。
修改评论