CORE--TRANSGENIC ANIMAL MODEL

核心--转基因动物模型

• 批准号: 6948013
• 负责人: Sally A. Camper
• 金额: $ 3万
• 依托单位: UNIVERSITY OF MICHIGAN AT ANN ARBOR
• 依托单位国家: 美国
• 财政年份: 2005
• 资助国家: 美国
• 起止时间: 2005-07-01 至 2010-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6948013
• 关键词: aging; animal colony; animal old age; biomedical facility; cryopreservation; electroporation; embryonic stem cell; gene targeting; genetically modified animals; germ free condition; laboratory mouse; laboratory rat; microinjections; mutant

This Core provides a complete set of services to the University of Michigan Nathan Shock Center (UM-NSC) scientists. These services include the production of transgenic mice, transgenic rats, and gene targeted ("knockout") mice from embryonic stem (ES) cells. Transgenic mice are used in many ways, including the tissue specific gene overexpression, production of animal models of human disease, and correction of disease phenotypes. 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 available services are rederivation of pathogen free mice and rats, and mouse embryo cryopreservation and recovery. The Core maintains specialized reagents for transgenic and gene targeting research. The availability of this Core obviates the need for Center members to invest in expensive equipment and specialized training in transgenic technology. Consultation on all aspects of transgenic and gene targeting research is provided, from the design of DNA constructs to gene expression analysis. Guaranteed Production of Transgenic Mice and Rats: We deliver an average of nine transgenic founder mice and nine founder rats per transgenic order. We 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 pluripotent mouse ES cells with gene targeting vectors, pick 480 ES cell clones, and provide DNA from the clones for genetic screening. The Core will expand gene targeted ES cell clones and prepare them for the blastocyst microinjection. Blastocyst Microinjection: The Core guarantees that at least 50 blastocysts will be microinjected to produce ES-cell mouse chimeras. Investigators will breed the chimeras for germline transmission. Commensurate with Center Member usage in the last five years, we project annual demand of 20 orders for transgenic mouse production, one order for transgenic rats, 6 orders for production of gene targeted ES cells, and 15 orders for blastocyst microinjections to make chimeras. In exchange for Center support, UM-NSC scientists will receive a 40% discount on the recharge rate for these services.

该核心为密歇根大学内森休克中心（UM-NSC）的科学家提供一整套服务。这些服务包括从胚胎干细胞生产转基因小鼠、转基因大鼠和基因靶向（“敲除”）小鼠。转基因小鼠的应用有很多，包括组织特异性基因过表达、人类疾病动物模型的建立和疾病表型的纠正。ES细胞技术用于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。其他可用的服务包括无病原体小鼠和大鼠的再衍生以及小鼠胚胎冷冻保存和恢复。核心保持转基因和基因靶向研究的专用试剂。有了这一核心，中心成员就不必投资昂贵的设备和转基因技术的专门培训。从DNA构建体的设计到基因表达分析，为转基因和基因靶向研究的各个方面提供咨询。保证生产转基因小鼠和大鼠：我们平均每个转基因订单提供9只转基因创始小鼠和9只创始大鼠。我们保证，每个提交给核心的DNA构建体至少会产生三个创始人。基因靶向ES细胞的生产：Core将用基因靶向载体电穿孔多能小鼠ES细胞，挑选480个ES细胞克隆，并从克隆中提供DNA用于遗传筛选。Core将扩增基因靶向的ES细胞克隆，并为囊胚显微注射做好准备。囊胚显微注射：核心保证至少50个囊胚将被显微注射，以产生ES细胞小鼠嵌合体。研究人员将培育嵌合体进行生殖系传播。与中心成员在过去五年的使用量相当，我们预计每年需要20个转基因小鼠生产订单，1个转基因大鼠订单，6个基因靶向ES细胞生产订单，15个囊胚显微注射订单，以制造嵌合体。作为对中心支持的交换，UM-NSC科学家将获得这些服务充值费率的40%折扣。