Gene Targeting & Transgenic Mouse

项目摘要（见说明）：基因靶向和转基因设施成立于20多年前，以协助AECC研究人员在人类疾病的转基因小鼠模型的产生。直到最近，基因靶向和转基因组件作为独立的共享资源运作，在CCSG的最后一次审查中，每个组件都被评为优秀。在搬迁到价格中心新的最先进的实验室空间后，这些设施被合并为一个基因靶向和转基因共享资源。该设施的转基因部分继续以高效率（~ 100%成功率）产生转基因小鼠品系，其通过将DNA序列（例如常规质粒载体或BAC克隆）通过原核注射或通过受精卵母细胞的慢病毒感染引入生殖系。对于每个项目，设施主管与个别研究人员进行咨询，并就转基因构建体设计提供建议，以及制作质粒和序列盒，以确保适合在小鼠中表达。通常有20-25名研究者使用这项服务，每年注射约100个结构。基因靶向组件继续通过在胚胎干细胞（ES）中使用基因靶向方法并将这些变化引入小鼠生殖系来提供小鼠基因组修饰服务。基因靶向服务包括产生常规敲除小鼠品系、敲入小鼠品系和具有条件等位基因的小鼠品系，用于基因的时间和空间消融。该装置获得基因靶向小鼠的成功率高达95%以上。最后，阿尔伯特·爱因斯坦医学院实施了一项新的基因修饰服务，为所有AECC研究人员提供快速和具有成本效益的转基因小鼠品系生产的完整服务。服务包括设计和生成简单和复杂的基因靶向载体，电穿孔和筛选各种遗传背景的胚胎干细胞系（129，B6和129/06），以及通过囊胚注射（由基因靶向设施进行）生成嵌合小鼠。基因修饰服务还开发了高通量产生靶向载体和筛选程序的新技术和程序，并于2012年初开始全面运作。

PROJECT SUMMARY (See instructions): The Gene Targeting and Transgenic facility was founded over 20 years ago to assist AECC researchers in the generation of genetically modified mouse models of human disease. Until recently, the Gene Targeting and Transgenic components operated as independent Shared Resources, each being rated outstanding at the last CCSG review. Following relocation to new state-of-the-art laboratory space in the Price Center, the facilities were merged into one Gene Targeting and Transgenic Shared Resource. The Transgenic component of the facility continues to generate, with high efficiency (-100% success rate), transgenic mouse strains through the introduction of DNA sequences, such as regular plasmid vectors or BAC clones into the germ line by pronuclear injection or by lentivirus infection of fertilized oocytes. For each project, the facility supervisor consults with individual investigators and advises on transgene construct design, as well as making plasmids and sequence cassettes available to ensure suitable for expression in the mouse. Typically 20-25 investigators use this service with approximately 100 constructs injected per year. The Gene Targeting component continues to provide services for modification of the mouse genome through the use of gene targeting methods in embryonic stem (ES) cells and introduction of these changes into the mouse germline. Gene Targeting services include the generation of conventional knockout mouse lines, knock-in mouse lines and mouse lines with conditional alleles for the temporal and spatial ablation of genes. The facility has a high success rate (>95%) for obtaining gene targeted mouse. Finally, a new Gene Modification Service was implemented by the Albert Einstein College of Medicine to provide a complete service for the rapid and cost efficient generation of genetically modified mouse lines to all AECC investigators. Services include the design and generation of simple and complex gene targeting vectors, electroporation and screening of embryonic stem cell lines of various genetic backgrounds (129, B6 and 129/06) and the generation of chimeric mice by blastocyst injection (performed by the Gene Targeting facility). The Gene Modification Service has also developed new technologies and procedures for high-throughput generation of targeting vectors and screening procedures and began full operation in early 2012.