Genome Editing Shared Resource

CORE 008 -基因组编辑共享资源 项目总结/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，是使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，以有效地生成，存储和共享基因改变的小鼠模型。GESR，其中 已经存在了25年，在过去的五年里发生了重大变化。五个优先 针对小鼠胚胎干细胞（mESCs）基因靶向的服务线变得不必要， CRISPR/Cas9的发现。因此，在2015年底，GESR将重点转向提高效率， 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 已经从97个CRISPR/Cas9实验中产生了突变小鼠，其技术成功率为100%。 过去的26个项目。GESR现在提供了一种全面的服务方法，其中资源执行项目 设计、注射前试剂验证、注射后对所产生幼仔的分析，以及第一个 这意味着可以生成一个新的基因组，以向研究者提供正确编辑的杂合小鼠。调查人员不再需要 广泛的分子生物学技能，甚至是CRISPR/Cas9基因编辑策略的知识。到 为了支持这些新的CRISPR生成的线的共享，GESR变得更加积极主动地鼓励 使用核心冷冻保存服务。小鼠品系的室内冷冻保存改善了集落 管理，并使特定的病原体免费股票作为冷冻保存的种质发送到其他 机构职能体系该资源继续重建VICC要求的冻存小鼠品系 来自其他研究者的研究者，或存在于外部储存库中的研究者， 受精和胚胎移植。最后，资源已经建立，并正在努力扩大，一个范德比尔特 冻存小鼠库（VCMR）。VCMR有助于遵守NIH鼠标共享政策 并且能够在没有研究者持续参与的情况下存储和分发来自范德比尔特的小鼠。 所有显微注射和胚胎移植服务都在无特定病原体屏障设施中进行。 总之，GESR提供的服务组合使VICC成员和其他范德比尔特 研究人员有效地生成和维护新的基因组编辑小鼠模型，以研究多个方面 细胞和肿瘤生物学的研究。

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.