Genome Editing Shared Resource

CORE 008-基因组编辑共享资源 项目摘要/摘要 基因组编辑共享资源(GESR)的使命，以前是转基因小鼠/胚胎 干细胞共享资源，是为了让范德比尔特-英格拉姆癌症中心(VICC)的研究社区， 包括我们的VICC成员和田纳西州立大学梅哈里医学院的合作伙伴以及其他 NCI癌症中心高效地生成、存储和共享转基因小鼠模型。Gesr，其中 已经存在了25年以上，在过去的五年里经历了重大变化。五个之前 在小鼠胚胎干细胞(MESCs)中以基因打靶为导向的服务线在 CRISPR/Cas9的发现。因此，在2015年末，Gesr将重点转向提高效率和 CRISPR/Cas9介导的基因编辑在单细胞和双细胞小鼠胚胎中的结果。从那时起，超过800人 已经从97个CRISPR/Cas9实验中产生了突变小鼠，技术成功率为100% 过去的26个项目。Gesr现在提供了一种资源执行项目的全方位服务方法 设计、注射前试剂验证、注射后分析所产生的幼崽，以及第一只幼崽的繁殖 为研究人员提供正确编辑的杂合子小鼠的一代。调查人员不再需要 广博的分子生物学技能，甚至更多关于CRISPR/Cas9基因编辑策略的知识。至 支持分享这些由CRISPR生成的新线路，GESR已变得更加积极主动地鼓励 使用核心冷冻保存服务。小鼠品系室内冷冻保存可改善菌落 管理并允许将特定的无病原菌的种质作为超低温保存的种质发送给其他 机构。该资源继续重建VICC要求的冷冻保存的小鼠品系 来自其他调查人员的调查人员，或存在于外部资料库中的调查人员，通过在体外执行这两种操作 受精和胚胎移植。最后，资源已经建立，并正在努力扩大，范德比尔特 冷冻保存小鼠库(VCMR)。VCMR有助于遵守NIH鼠标共享政策 并使范德比尔特的老鼠能够在不需要调查人员继续参与的情况下储存和分发。 所有显微注射和胚胎移植服务都是在特定的无病原体屏障设施中进行的。 总而言之，GESR提供的服务组合使VICC成员和其他Vanderbilt 研究人员高效地生成和维护新的基因组编辑小鼠模型，以研究多方面 使用复杂的小鼠模型研究细胞和肿瘤生物学。

CORE 008 – GENOME EDITING SHARED RESOURCE PROJECT SUMMARY/ABSTRACT The mission of the Genome Editing Shared Resource (GESR), previously the Transgenic Mouse/Embryonic Stem Cell Shared Resource, is to enable the Vanderbilt-Ingram Cancer Center (VICC) research community, inclusive of our VICC members and partners at Meharry Medical College, Tennessee State University, and other NCI Cancer Centers to efficiently generate, store and share genetically-altered mouse models. GESR, which has been in existence for over 25 years, has undergone major changes over the past five years. Five prior service lines oriented around gene targeting in mouse embryonic stem cells (mESCs) became unnecessary after the discovery of CRISPR/Cas9. Thus, in late 2015, GESR shifted its focus towards improving the efficiency and outcomes of CRISPR/Cas9-mediated gene editing in one and two cell mouse embryos. Since then, over 800 mutant mice from 97 CRISPR/Cas9 experiments have been generated, with a 100% technical success rate for the past twenty-six projects. GESR now provides a full-service approach where the resource performs project design, pre-injection reagent validation, post-injection analysis of the resulting pups, and breeding of the first generation to provide the investigator with a correctly edited heterozygous mouse. Investigators no longer need extensive molecular biology skills, or even much knowledge about CRISPR/Cas9 gene editing strategies. To support the sharing of these new CRISPR-generated lines, GESR has become more proactive in encouraging the use of core cryopreservation services. In house cryopreservation of mouse lines improves colony management and enables specific pathogen free stocks to be sent as cryopreserved germplasm to other institutions. The resource continues to reconstitute cryopreserved mouse lines that are requested by VICC investigators from other investigators, or that exist within external repositories, by performing both in vitro fertilization and embryo transfer. Finally, the resource has established, and are striving to expand, a Vanderbilt Cryopreserved Mouse Repository (VCMR). The VCMR facilitates compliance with NIH Mouse Sharing Policies and enables storage and distribution of mice from Vanderbilt without the continued involvement of investigators. All microinjection and embryo transfer services are performed in a specific pathogen free barrier facility. Together, the combination of services provided by the GESR enables VICC members and other Vanderbilt investigators to efficiently generate and maintain novel genome-edited mouse models to study multiple aspects of cell and tumor biology using sophisticated mouse models.