Transgenic Core

转基因核心公司与研究人员合作生成动物模型，这将增加我们的 对风湿性疾病基因功能的基本认识。这一核心成立于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(敲除) 会员。其他服务包括无病原体小鼠的再衍生、小鼠品系的冷冻保存和 康复和转基因技术培训。转基因核心维护着专门的设备 显微注射、冷冻保存和小鼠胚胎干细胞培养。核心维护和 为构建转基因或基因打靶载体分发质粒。最大限度地提高基因靶向性 成功，Core对ES细胞系、饲养层细胞和ES细胞血清进行质量保证测试 文化。这是一个合作的核心，结合了研究人员在分子生物学方面的专业知识 他们研究的基因和Core在培育基因工程小鼠方面的专业知识。 使这一核心脱颖而出的独特能力是1)转基因小鼠和大鼠的保证生产，2) 常规生产BAG转基因小鼠，3)在独特遗传背景下生产转基因小鼠， 4)在除129/Sv ES细胞外的C57BL/6 ES细胞系中进行基因打靶，5)开放试剂和 设备，以及胚胎干细胞培养和显微注射方法方面的培训。获得转基因核心 消除了调查人员将资源投入设备采购和人员时间 在显微操作、胚胎干细胞培养和小鼠胚胎操作方面的培训。各方面的协商 从转基因设计和条件靶向，提供了转基因和ES细胞研究的最新进展 载体应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名创始小鼠 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以进行筛选 与靶向载体进行同源重组。我们保证用所需的基因克隆ES细胞 这些改变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。这个 这些程序的效率达到或超过了文献中公布的值。 根据过去的使用情况，中心成员预计每年使用30只转基因小鼠，6只ES 细胞电穿孔和15个ES细胞-小鼠嵌合体。这是大学核心学院总人数的34% 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.