Transgenic Core

转基因核心

• 批准号: 7483083
• 负责人: Sally A. Camper
• 金额: $ 7.38万
• 依托单位: UNIVERSITY OF MICHIGAN AT ANN ARBOR
• 依托单位国家: 美国
• 财政年份: 2007
• 资助国家: 美国
• 起止时间: 2007-08-01 至 2011-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7483083
• 关键词: Animal Model; Artificial Chromosomes; BAC (bacterial artificial chromosome); Bacterial Artificial Chromosomes; Bos taurus; Breeding; Cattle; Cells; Chimera organism; Chromosomes; Chromosomes, Artificial, Yeast; Consultations; Cosmids; Count; Cryopreservation; Cultured Cells; Custom; DNA; ES Cell Line; Electroporation; Embryo; Embryo Transfer; Ensure; Equipment; Facility Construction Funding Category; Fostering; Freezing; Gene Targeting; Genes; Genetic; Genetically Engineered Mouse; Genomics; Genotype; Hand; Human Resources; Inbred F344 Rats; Internet; Knockout Mice; Knowledge; Laboratory culture; Liquid substance; Literature; Methods; Microinjections; Micromanipulation; Modification; Molecular Biology; Monitor; Mothers; Mouse Strains; Mouse, Founder, Transgenic; Mus; Mutant Strains Mice; Mutation; Nitrogen; P1 Bacteriophage Artificial Chromosomes; Paper; Phenotype; Philosophy; Plasmids; Procedures; Production; Publishing; Quality Control; Rattus; Reagent; Recovery; Research; Research Personnel; Resources; Review Literature; Rheumatism; SWR/J Mouse; Serum; Services; Site; Source; Stem Cell Research; Technology; Testing; Time; Training; Transgenes; Transgenic Animals; Transgenic Mice; Transgenic Organisms; Weaning; Week; Work; base; blastocyst; cost; day; design; desire; egg; embryonic stem cell; fetal; gene function; germ free condition; homologous recombination; innovation; instrumentation; interest; member; new technology; pathogen; pup; quality assurance; stem; success; transmission process; vector; zygote

The Transgenic Core works with investigators to generate animal models that will increase our fundamental understanding of gene function in rheumatic diseases. This Core was established in 1989 and produces transgenic mice and rats, and gene-targeted mice (knockouts) for Rheumatic Disease Core Center members. Other services include rederivation of pathogen free mice, mouse strain cryopreservation and recovery, and transgenic technology training. The Transgenic Core maintains specialized equipment for microinjection, cryopreservation, and mouse embryonic stem (ES) cell culture. The Core maintains and distributes plasmids for transgene or gene targeting vector construction. To maximize gene targeting success, the Core performs quality assurance tests on ES cell lines, feeder cells, and serum for ES cell culture. This is a collaborative Core that combines the expertise of investigators in the molecular biology of the genes they study and the Core's expertise in producing genetically engineered mice. Unique capabilities that set this Core apart are 1) guaranteed production of transgenic mice and rats, 2) routine production of BAG transgenic mice, 3) production of transgenic mice in unique genetic backgrounds, 4) gene targeting in C57BL/6 ES cell lines in addition to 129/Sv ES cells, 5) open access to reagents and equipment, and training in ES cell culture and microinjection methods. Access to the Transgenic Core obviates the need for investigators to devote resources to equipment purchases and personnel time to training in micromanipulation, ES cell culture, and mouse embryo manipulation. Consultation on all aspects of transgenic and ES cell research is provided, from the design of transgenes and conditional targeting vectors to mouse breeding and phenotype analysis. We deliver an average of ten transgenic founder mice and guarantee that at least three founders will be produced for each DMA construct submitted to the Core. The Core electroporates totipotent ES cells with targeting vectors, selects 480 ES cell clones, and provides investigators with ES cell clone DNA to screen for homologous recombination with targeting vectors. We guarantee that ES cell clones with desired genetic changes will be microinjected into at least 50 mouse blastocysts to produce ES cell-mouse chimeras. The efficiency of these procedures meets or exceeds published values in the literature. Based on past use, the projected annual usage by Center members is 30 transgenic mouse orders, 6 ES cell electroporation orders, and 15 ES cell-mouse chimera orders. This is 34% of the Core's total universitywide capacity and is consistent with past usage by Center Members.

转基因核心与研究人员合作，生成动物模型，以增加我们的 对风湿病中基因功能的基本理解。该核心成立于1989年， 产生转基因小鼠和大鼠，以及风湿病核心中心的基因靶向小鼠（敲除） 成员。其他服务包括无病原体小鼠的重新培训，小鼠菌株冷冻保存和 恢复和转基因技术培训。转基因核心维护专业设备 显微注射，冷冻保存和小鼠胚胎（ES）细胞培养。核心保持和 分布用于转基因或靶向载体构建基因的质粒。最大化基因靶向 成功，核心对ES细胞系，进料细胞和ES细胞的血清进行质量保证测试 文化。这是一个协作核心，结合了研究人​​员在分子生物学中的专业知识 他们研究的基因以及核心在产生基因工程小鼠方面的专业知识。 设置此核心的独特功能是1）保证产生转基因小鼠和大鼠，2） 袋子转基因小鼠的常规产生，3）在独特的遗传背景中生产转基因小鼠， 4）除129/SV ES细胞外，C57BL/6 ES细胞系中的基因靶向，5）开放访问试剂和 设备和ES细胞培养和微注射方法的培训。访问转基因核心 消除了调查人员将资源投入设备购买和人员时间的需求 微观渗透，ES细胞培养和小鼠胚胎操作中的训练。有关各个方面的咨询 从转基因和条件靶向的设计提供了转基因和ES细胞研究 小鼠育种和表型分析的向量。 我们平均提供十只转基因创始人小鼠，并保证至少三个创始人将是 为提交给核心的每个DMA构造生产。核心电孔与 靶向向量，选择480 ES细胞克隆，并为研究人员提供ES Clone DNA筛选 靶向向量的同源重组。我们保证具有所需遗传的ES细胞克隆 变化将被微注射到至少50个小鼠胚泡中，以产生ES细胞鼠嵌合体。这 这些程序的效率符合或超过文献中公布的价值。 根据过去的使用，中心成员预计的年度使用是30个转基因鼠标订单，6 es 细胞电穿孔顺序和15个ES细胞鼠嵌合订单。这是核心总范围内的34％ 能力，与中心成员过去的使用一致。