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，这将提高我们的 对风湿病基因功能的基本了解。该核心成立于 1989 年， 为风湿病核心中心生产转基因小鼠和大鼠以及基因靶向小鼠（基因敲除） 成员。其他服务包括无病原体小鼠的再衍生、小鼠品系冷冻保存和 恢复和转基因技术培训。转基因核心拥有专门的设备 显微注射、冷冻保存和小鼠胚胎干 (ES) 细胞培养。核心维护并 分发用于转基因或基因靶向载体构建的质粒。最大化基因靶向 成功后，Core 对 ES 细胞系、饲养细胞和 ES 细胞血清进行质量保证测试 文化。这是一个协作核心，结合了分子生物学研究人员的专业知识 他们研究的基因以及核心在生产基因工程小鼠方面的专业知识。 使该核心与众不同的独特功能是 1) 保证转基因小鼠和大鼠的生产，2) BAG转基因小鼠的常规生产，3）独特遗传背景下转基因小鼠的生产， 4) 除 129/Sv ES 细胞外，还可在 C57BL/6 ES 细胞系中进行基因靶向，5) 开放试剂和 设备以及 ES 细胞培养和显微注射方法的培训。获得转基因核心 调查人员无需投入资源购买设备和人员时间 显微操作、胚胎干细胞培养和小鼠胚胎操作方面的培训。各方面咨询 从转基因设计和条件靶向提供转基因和胚胎干细胞研究 小鼠育种和表型分析的载体。 我们平均提供十只转基因创始人小鼠，并保证至少三名创始人将被 为提交给核心的每个 DMA 构造生成。 Core 电穿孔全能 ES 细胞 靶向载体，选择480个ES细胞克隆，并为研究人员提供ES细胞克隆DNA进行筛选 与靶向载体同源重组。我们保证 ES 细胞克隆具有所需的遗传 这些变化将被显微注射到至少 50 个小鼠囊胚中，以产生 ES 细胞-小鼠嵌合体。这 这些程序的效率达到或超过文献中公布的值。 根据过去的使用情况，中心成员预计每年的使用量为 30 只转基因小鼠订单、6 只 ES 细胞电穿孔订单和 15 个 ES 细胞-小鼠嵌合订单。这是整个大学核心核心总数的 34% 容量并与中心成员过去的使用情况一致。