Transgenic Core

转基因核心与研究人员合作，产生动物模型，将增加我们的 对风湿性疾病中基因功能的基本认识。该中心成立于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（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 康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。核心保持和 分发用于转基因或基因靶向载体构建的质粒。为了最大化基因靶向 成功后，该中心对ES细胞系、饲养细胞和ES细胞血清进行了质量保证试验 文化这是一个合作的核心，结合了研究人员在分子生物学的专业知识， 他们所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。 使这个核心与众不同的独特能力是：1）保证生产转基因小鼠和大鼠，2） 常规生产BAG转基因小鼠，3）生产独特遗传背景的转基因小鼠， 4)除129/Sv ES细胞外，还在C57 BL/6 ES细胞系中进行基因靶向，5）开放获得试剂， 胚胎干细胞培养和显微注射方法的培训。进入转基因核心 使调查人员无需投入资源购买设备和人员时间， 显微操作、ES细胞培养和小鼠胚胎操作培训。各方面的协商 从转基因设计和条件靶向两个方面介绍了转基因和ES细胞研究的最新进展 载体用于小鼠育种和表型分析。 我们平均提供10只转基因创始小鼠，并保证至少有3只创始小鼠将被 为提交给核心的每个DMA构建体生成。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以筛选 与靶向载体的同源重组。我们保证具有所需基因的ES细胞克隆 将这些变化显微注射到至少50个小鼠胚泡中以产生ES细胞-小鼠嵌合体。的 这些方法的效率达到或超过文献中公开的值。 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 细胞电穿孔顺序和15个ES细胞-小鼠嵌合体顺序。这是34%的核心的总全校 容量，并与中心成员过去的使用情况一致。

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.