遗传 ›› 2021, Vol. 43 ›› Issue (7): 704-714.doi: 10.16288/j.yczz.21-163

• 资源与平台 • 上一篇    

全基因组蛋白质的标签细胞和小鼠资源库建设

蒋婧(), 赵安麒, 谢甜, 陈淑藯, 李劲松   

  1. 中国科学院分子细胞科学卓越创新中心(生物化学与细胞生物学研究所), 基因组标签计划研发中心,上海 200031
  • 收稿日期:2021-04-29 修回日期:2021-06-07 出版日期:2021-07-20 发布日期:2021-06-24
  • 通讯作者: 蒋婧 E-mail:jiangjing@sibcb.ac.cn
  • 基金资助:
    上海市科学技术委员会科技创新行动计划项目编号(17411954900);上海市科学技术委员会科技创新行动计划项目编号(19411951800);中国科学院战略性先导科技专项编号(XDB19010204);国家自然科学基金项目资助编号(31801057)

Construction of genome-wide protein tagging cell and mouse libraries

Jing Jiang(), Anqi Zhao, Tian Xie, Shuwei Chen, Jinsong Li   

  1. Genome Tagging Project Center, Center for Excellence in Molecular Cell Science, Shanghai Institute of Biochemistry and Cell Biology, Chinese Academy of Sciences, Shanghai 200031, China
  • Received:2021-04-29 Revised:2021-06-07 Online:2021-07-20 Published:2021-06-24
  • Contact: Jiang Jing E-mail:jiangjing@sibcb.ac.cn
  • Supported by:
    Supported by Shanghai Municipal Commission for Science and Technology Grants Nos(17411954900);Supported by Shanghai Municipal Commission for Science and Technology Grants Nos(19411951800);the Strategic Priority Research Program of Chinese Academy of Sciences No(XDB19010204);the National Natural Science Foundation of China No(31801057)

摘要:

小鼠是最广泛使用的模式生物,构建基因改造的小鼠模型是研究基因功能和疾病发生机制的重要手段。从20世纪80年代发展至今,小鼠基因改造技术更新迭代,主要包括胚胎干细胞介导的同源重组策略、配子介导的转基因策略、以及最新基于CRISPR/Cas9技术的遗传改造策略等。由我国科学家自主研发的全新小鼠基因改造技术——“人造精子细胞”(或“类精子干细胞”)介导的半克隆技术与以往策略相比,具有小鼠构建周期短、效率高、成本低、应用场景兼容性强等特点。为了摆脱我国小鼠实验动物“卡脖子”风险,填补标签蛋白质小鼠资源库的国际空白,中国科学院分子细胞科学卓越创新中心进行了战略资源布局,于2017年启动了基因组标签计划(genome tagging project, GTP),并成立了GTP研发中心致力于建立全新的标签细胞和小鼠战略资源库。GTP研发中心通过对“人造精子细胞”基因组上编码蛋白质的基因原位系统性地添加标签序列,建立了可用液氮保存的标签细胞资源库,并作为生产标签小鼠的种子贮备。当需要标签小鼠时,利用半克隆技术将标签细胞注射到小鼠卵子中,一步法获得标签小鼠。标签小鼠可通过标准化的标签抗体监测特定蛋白质从而实现蛋白质的在体、实时、动态研究。目前,GTP研发中心已构建1532个标签细胞和277个标签小鼠,初步形成了具有特色标签产品战略资源库;已有242个标签小鼠品系被分发到3个囯家15个地区的32个研究机构,供66个研究团队使用。同时,GTP研发中心已建立完备的标签产品信息,定期在GTP官网上发布更新,供科研人员查询、订购和定制。后续标签小鼠的配繁规律、标签蛋白质组织图谱描绘、发表文献等也会陆续更新到GTP官网。GTP研发中心将对标行业尖端,打造全球独特性战略资源,为蛋白质功能研究提供标准化平台,推动生命科学的发展和临床转化。

关键词: 基因组标签计划, “人造精子细胞”, 半克隆技术, 标签小鼠, 战略资源

Abstract:

Mice are the most widely used model organism for the study of gene functions and disease mechanisms through the generation of gene-modified mice. Since the 1980s, different genetic manipulation technologies have been developed to reveal gene functions in vivo, including homologous recombination strategies mediated by embryonic stem cells, transgenic strategies mediated by gametes, and the latest genetic modification strategies based on CRISPR/Cas9 technology. Semi-cloning technology mediated by “artificial spermatids” (androgenetic haploid embryonic stem cells, also termed sperm-like stem cells) is developed by Chinese scientists in 2012. In combination with CRISPR/Cas9, semi-cloning technology enables one-step generation of gene-modified mice through injection of “artificial spermatids” with specific gene modifications into oocytes. It has the characteristics of short construction cycle, high efficiency, low cost, and high application compatibility. In 2017, the Center for Excellence in Molecular Cell Science (CEMCS) of CAS has launched the genome tagging project (GTP) based on “artificial spermatid”-mediated semi-cloning technology. The ambitious goal of GTP is to tag every protein in mice and construct a unique mouse library that maintains the genome-wide protein-tagging mouse models. Subsequently, the GTP center was established at CEMCS to pursue the project. GTP center developed strategies to generate protein-tagging cells and mice. Briefly, a tag sequence is precisely inserted in a specific protein- coding gene endogenously in cultured “artificial spermatids”in vitro to build a cell library, in which, each cell line carrying a specific protein tag. The tagged cells could be further used as a sperm replacement to produce tagged mice in one step upon injection into oocytes. The tagged mouse library enables global analysis of protein expression, localization, and complexes using standard tag-based assays in vivo. By April 2021, the GTP center has generated 1532 tagged cell lines, 277 of which have been successfully used to produce tagged mice through oocyte injection. A total of 242 tagged mouse strains have been distributed to 66 research teams in 32 research institutions of 15 districts in 3 countries. The database of tagging product resources has been established and released regularly on the GTP website for scientists to inquire and order. Later, more information about GTP products, such as mouse breeding, protein tissue expression map, published literature, etc., will also be successively published on the GTP website. The GTP center will provide a standardized platform for protein function research, which may dramatically promote the development of life science and clinical transformation.

Key words: genome tagging project, artificial spermatid, semi-cloning technology, protein tagging mouse, strategic resource