GENE TARGETING SHARED RESOURCE

基因靶向共享资源

• 批准号: 7506851
• 负责人: WINFRIED EDELMANN
• 金额: $ 12.33万
• 依托单位: ALBERT EINSTEIN COLLEGE OF MEDICINE
• 依托单位国家: 美国
• 财政年份: 2007
• 资助国家: 美国
• 起止时间: 2007-09-25 至 2012-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7506851
• 关键词: Ablation; Albert Einstein Cancer Center; Animal Model; Animals; Blastocyst Transfer; Cancer Center; Cells; Chimera organism; Cloning; Collaborations; Communities; Complex; Country; ES Cell Line; Embryo; Facility Construction Funding Category; Faculty; Female; Fibroblasts; Funding; Future; Gene Targeting; Generations; Genes; Genetic; Genetic Recombination; Genomic Library; Genomics; Goals; Human; Induced Mutation; Injection of therapeutic agent; Institution; Knock-in Mouse; Knock-out; Malignant Neoplasms; Methodology; Methods; Microinjections; Modeling; Modification; Mouse Models of Human Cancer Consortium; Mus; NCI-Designated Cancer Center; Partner in relationship; Price; Procedures; Process; Production; Rate; Research Personnel; Resistance; Resource Sharing; Services; Source; Techniques; Technology; Training; United States National Institutes of Health; base; blastocyst; design; developmental genetics; embryonic stem cell; human cancer mouse model; human disease; male; member; mouse genome; mouse model; new technology; repository; stem; success; translational medicine; transmission process; vector

The Gene Targeting Facility was established in 1992 to assist AECC researchers in the generation of animal models for human cancer. The goals of the facility are to provide services that allow researchers to modify the mouse genome through the methods of gene targeting in embryonic stem (ES) cells and to introduce these changes into the mouse germline. The facility provides: (1) Advice in the theoretical and practical aspects of gene targeting vector design including design of conventional knockout targeting vectors, knock-in targeting vectors as well as targeting vectors for temporal and spatial ablation of genes. The facility also advises on the methodologies required for targeting vector construction, including the use of gridded BAG 129 strain genomic libraries for the rapid isolation of genomic fragments, subcloning strategies and also the use of recombination based cloning. The facility maintains and makes available a panel of vectors and resistance cassettes that facilitate targeting vector construction. In addition, the facility established recombination-based cloning technology ("recombineering") using the RecET technology that allows the modification of BAG clones and also permits the more rapid generation of more complex targeting vectors. (2) The facility provides comprehensive ES cell culture service and maintains stocks of mouse embryonic fibroblasts (MEF) for feeder cell generation and ES cell lines with high germline transmission capacity. The facility offers complete ES cell culture service encompassing all the required techniques but will train and provide advice to investigators who wish to participate in parts of this process. The facility provides services for chimera production. This includes ES cell microinjection into blastocysts, blastocyst transfer and chimera production. To support these activities, stocks of ES cell lines from different sources and of different genetic backgrounds (129J/Sv, 129/B6 and B6) are maintained along with core mouse colonies sufficient to generate enough blastocysts for injection, as well as pseudopregnant recipient mice for the blastocyst transfers and vasectomized males to mate with these females. In addition, the facility is intimately involved in the implementation of new technologies and the training of new investigators in the technical and theoretical aspects of these procedures. This facility will be relocated to the new Price Center for Genetic and Translational Medicine in a larger space adjacent to a new Animal Barrier Shared Resource when that facility opens in December 2007.

基因打靶设施成立于 1992 年，旨在协助 AECC 研究人员培育动物 人类癌症模型。该设施的目标是提供允许研究人员修改的服务 通过胚胎干（ES）细胞中的基因打靶方法对小鼠基因组进行改造，并引入 这些变化进入小鼠种系。该设施提供： (1) 理论和实践方面的建议 基因靶向载体设计的各个方面，包括常规敲除靶向载体的设计， 敲入靶向载体以及用于基因时间和空间消融的靶向载体。设施 还就靶向载体构建所需的方法提供建议，包括网格的使用 BAG 129 菌株基因组文库，用于快速分离基因组片段、亚克隆策略以及 使用基于重组的克隆。该设施维护并提供一组载体和 促进靶向载体构建的抗性盒。此外，该设施还设立了 基于重组的克隆技术（“重组工程”），使用 RecET 技术，允许 BAG 克隆的修饰还允许更快地生成更复杂的靶向载体。 （2）该设施提供全面的ES细胞培养服务并维持小鼠胚胎库存 用于饲养细胞生成的成纤维细胞（MEF）和具有高种系传递能力的 ES 细胞系。这 设施提供完整的 ES 细胞培养服务，涵盖所有必需的技术，但将培训和 向希望参与此过程部分的调查人员提供建议。该设施提供服务 用于嵌合体生产。这包括将 ES 细胞显微注射到囊胚中、囊胚移植和 嵌合体生产。为了支持这些活动，来自不同来源和不同类型的 ES 细胞系库存 遗传背景（129J/Sv、129/B6 和 B6）与核心小鼠群体一起维持，足以 产生足够的囊胚用于注射，以及产生囊胚的假孕受体小鼠 转移并切除输精管的雄性与这些雌性交配。此外，该设施还密切参与 新技术的实施以及技术和新研究人员的培训 这些程序的理论方面。该设施将搬迁至新的遗传价格中心 和转化医学在一个更大的空间毗邻新的动物屏障共享资源 设施于 2007 年 12 月开业。