CORE--TRANSGENIC FACILITY

转基因动物模型核心从携带有针对性基因改变的胚胎干细胞中培育出转基因小鼠和基因靶向(“基因敲除”)小鼠。转基因小鼠被用于许多方面，包括研究组织特异性基因表达，制作人类疾病的动物模型，以及测试用于基因治疗的基因构建的有效性。ES细胞技术被用来建立隐性遗传病的动物模型，并表征基因丢失和基因细微变化的功能效应。ES细胞技术被用来建立隐性遗传病的动物模型，并表征基因丢失和基因细微变化的功能效应。提供的其他服务包括小鼠胚胎的冷冻保存和复苏，无病原体小鼠的衍生，以及咨询。Core拥有专门的转基因和基因靶向研究试剂。核心的专门设备用于支持多名调查人员的研究工作，否则这些调查人员无法获得此类设备。现场提供转基因核心使研究人员不需要购买昂贵的设备并投入人员时间进行长期的微操作技术培训，这对大多数研究人员来说在技术上和经济上都是不可能的。提供转基因和胚胎干细胞研究的所有方面的咨询，从DNA结构的设计到小鼠饲养。转基因小鼠的生产：我们平均提供10只转基因创始小鼠，并保证每个提交给Core的DNA构建将产生至少三个创始小鼠。基因靶向ES细胞的生产：Core将用Tar4getinbg载体电穿孔全能ES细胞，选择ES细胞克隆，并向中心成员提供克隆的DNA用于基因筛选。ES细胞-小鼠嵌合体的生产：携带所需基因突变的ES细胞克隆将被显微注射到小鼠囊胚中，这些囊胚将通过手术转移到养母体内以产生嵌合体。转基因技术培训：研究人员还可以选择接受培训，1)通过原核显微注射产生转基因小鼠，2)操纵ES细胞并产生基因靶向的ES细胞克隆，3)通过胚泡注射产生ES细胞-小鼠嵌合体。根据过去四年中心成员的使用情况，我们预计每年对转基因小鼠的需求为20个订单，生产基因靶向ES细胞的订单为6个订单，ES细胞-小鼠嵌合体的订单为20个订单。中心会员将获得这些服务的充值率35%的折扣。转基因核心在过去四年中完成的所有工作中，有40%致力于中心成员。

The Transgenic Animal Model Core produces transgenic mice and gene targeted ("knockout") mice from embryonic stem (ES) cells carrying targeted genetic alterations. Transgenic mice are used in many ways, including the study of tissue specific gene expression, production of animal models of human disease, and testing efficacy of gene constructs for gene therapy. ES cell technology is used to establish animal models of recessive genetic disease and to characterize functional effects of gene loss and subtle changes in genes. ES cell technology is used to establish animal models of recessive genetic disease and to characterize functional effects of gene loss and subtle changes in genes. Other services provided include mouse embryo cryopreservation and recovery, derivation of pathogen free mice, and consultation. The Core maintains specialized reagents for transgenic and gene targeting research. The Core's specialized equipment is used to support the research efforts of multiple investigators who otherwise lack access to such equipment. The availability of a transgenic Core on site obviates the need for individual researchers to purchase costly equipment and invest personnel time in lengthy training in micromanipulation techniques, which would be technically and financially impossible for most investigators. Consultation on all aspects of transgenic and ES cell research is provided, from the design of DNA constructs to mouse husbandry. Production of Transgenic Mice: We deliver an average of ten transgenic founder mice and guarantee that at least three founders will be produced for each DNA construct submitted to the Core. Production of Gene Targeted ES Cells: The Core will electroporate totipotent ES cells with tar4getinbg vectors, select ES cell clones, and provide Center Members with DNA from the clones for genetic screening. Production of ES Cell-Mouse Chimeras: ES cell clones carrying the desired genetic mutation will be microinjected into mouse blastocysts which are surgically transferred to foster mothers to produce chimeras. Transgenic Technology Training: Researchers can also elect to be trained to 1) produce transgenic mice by pronuclear microinjection, 2) manipulate ES cells and produce gene targeted ES cell clones, and 3) produce ES cell-mouse chimeras by blastocyst injection. Commensurate with Center Member usage in the last four years, we project annual demand for 20 orders for transgenic mice, 6 orders for production of gene targeted ES cells, and 20 orders for ES cell-mouse chimeras. Center members will receive a 35% discount on the recharge rate for these services. Forty percent of all work done in the last four years by the Transgenic Core was devoted to Center members.