Mouse Genetics

摘要 小鼠遗传学核心的主要作用是促进在MSK使用小鼠分子遗传学， 与癌症密切相关的基因功能的体内研究。MSK研究人员发展的相关领域， 应用小鼠模型包括：细胞生长和行为，干细胞生物学，胚胎发育， 免疫生物学、基因组完整性、癌症生物学和实验治疗学。MGC由2个 转基因/胚胎学组（TgG）和分子生物学/组织培养组（MBTCG）。 TgG提供的主要服务包括通过原核技术生产基因工程小鼠（GEM）。 注射DNA或CRISPR/Cas9和通过胚泡注射ES细胞;冷冻保存和长期 GEM系的储存;通过IVF和/或胚胎移植进行菌株再衍生或恢复; 特殊胚胎学和动物外科手术;以及提供GEM系，如CRE和FLPe 表达菌株。MBTCG提供的专业分子生物学服务包括转基因 建立者小鼠的DNA纯化和基因分型; CRISPR/Cas9试剂的设计和生产 小鼠基因组编辑;以及基因编辑小鼠携带的等位基因的鉴定和验证。服务 MBTCG提供的技术还包括在小鼠ES细胞中进行基因靶向的所有方面， 从GEM系建立ES细胞。MGC的一个组成部分是提供咨询和培训， 小鼠模型的设计和生产、ES细胞培养以及小鼠饲养和遗传学。发展也很快 在基因组编辑和测序技术中，结合翻译的稳定增加， MSK的研究，导致对MGC服务的需求大幅增加。另外核心 继续追求和实施新的技术发展，这将加快小鼠的基因编辑; 正在进行的努力包括使用商业上可获得的合成gRNA组分和应用 电穿孔以将CRIPSR/Cas9递送到小鼠受精卵中。将这些进步融入核心的 服务无疑将提高生产率，并减少员工为MSK创建GEM模型所需的时间/精力 investigators.

ABSTRACT The primary role of the Mouse Genetics Core is to facilitate the use of mouse molecular genetics at MSK for in vivo studies of gene functions germane to cancer. Relevant fields in which MSK investigators develop and apply mouse models include: cell growth and behavior, stem cell biology, embryonic development, immunobiology, genome integrity, cancer biology, and experimental therapeutics. The MGC consists of 2 sections: the Transgenic/Embryology Group (TgG) and the Molecular Biology/Tissue Culture Group (MBTCG). Major services offered by the TgG include the production of genetically engineered mice (GEM) by pronuclear injection of DNA or CRISPR/Cas9 and by blastocyst injection of ES cells; cryopreservation and the long-term storage of GEM lines; strain rederivation or recovery through IVF and/or embryo transfer; performance of special embryological and animal surgical procedures; and the provision of GEM lines such as CRE and FLPe expressing strains. The specialized molecular biology services provided by the MBTCG include transgene DNA purification and genotyping of founder mice; the design and production of CRISPR/Cas9 reagents for mouse genome editing; and the identification and verification of alleles carried by gene edited mice. Services offered by the MBTCG also include performing all aspects of gene targeting in mouse ES cells and establishing ES cells from GEM lines. An integral role of the MGC is to provide consultation and training in the design and production of mouse models, ES cell culture, and mouse husbandry and genetics. Rapid advances in genome editing and sequencing technologies, in combination with the steady increase in translational research at MSK, have resulted in a significant upsurge in demand for MGC services. In addition, the Core continues to pursue and implement new technical developments that will expedite gene editing in mice; ongoing efforts include the use of commercially available synthetic gRNA components and the application of electroporation to deliver CRIPSR/Cas9 into mouse zygotes. Integrating such advancements into the Core’s services will undoubtedly increase the productivity and reduce staff time/effort to create GEM models for MSK investigators.