Gene Targeting

项目总结（见说明）： 基因靶向共享资源成立于10年前，旨在帮助癌症中心成员使用动物模型构建和管理转基因小鼠品系。位于伍德伯里动物共享资源，该共享资源提供的服务包括通过原核注射DNA产生转基因小鼠，小鼠胚胎干（ES）细胞中的基因靶向，用遗传改变的ES细胞产生ES细胞嵌合体，以及为两者提供冷冻保存服务。 胚胎和精子。为了进一步扩大这些服务，该设施最近开发了通过四倍体囊胚注射生产ES细胞衍生小鼠的方案，这一程序极大地促进了CSHL癌症小鼠模型的快速生成。它还产生了用于C57 BL/6 J小鼠基因靶向的ES细胞系，当完全整合时，将使研究人员能够产生纯小鼠品系 在最广泛使用的遗传背景之一迅速。最后，基因靶向共享资源增加了核型分析作为嵌合小鼠生产的一个步骤;通过鉴定具有正常染色体互补的ES细胞，生殖系传播的效率大大提高。资源中新技术的快速整合有助于将癌症中心的研究保持在 尖端技术此外，其内部开发工作继续为癌症中心的研究人员提供新的工具和方法，以加快小鼠模型的生产，同时保持低成本。

PROJECT SUMMARY (See Instructions): The Gene Targeting Shared Resource was established 10 years ago to assist Cancer Center members who use animal models in constructing and managing genetically altered mouse strains. Located in the Woodbury Animal Shared Resource, this Shared Resource provides services including the generation of transgenic mice by pronuclear injection of DNA, gene targeting in mouse embryonic stem (ES) cells, generation of ES cell chimeras with genetically altered ES cells, and cryopreservation services for both embryos and sperm. To expand on these services further, the facility has recently developed protocols for producing ES cell-derived mice by tetraploid blastocyst injections, a procedure that has greatly facilitated the rapid generation of cancer mouse models at CSHL. It also generated ES cell lines for gene targeting in C57BL/6J mice, which, when fully incorporated, will enable researchers to produce pure mouse strains rapidly in one of the most widely used genetic backgrounds. Finally the Gene Targeting Shared Resource has added karyotyping as a step in the production of chimeric mice; by identifying ES cells with a normal chromosome complement, the efficiency of germline transmission has been greatly enhanced. The rapid incorporation of new technologies within the Resource helps keep research in the Cancer Center at the cutting edge. In addition, its internal development efforts continue to provide Cancer Center investigators with new tools and method to speed up the production of mouse models while maintaining low costs.