CORE--TRANSGENIC MOUSE/ES CELL CHIMERA FACILITY

转基因小鼠/胚胎干细胞嵌合体设施提供四种基本 服务：通过注射受精卵生产转基因(TG)小鼠， 胚胎干细胞嵌合动物的生产，再衍生 小鼠品系及其基因靶向ES细胞的分离 同源重组。为了生产转基因小鼠，用户 向受精卵提供适合于亲核注射的DNA构建物 老鼠蛋。转基因小鼠是在远缘繁殖的瑞士韦伯斯特小鼠中产生的， 近交系FVB/N小鼠或特定基因的小鼠。潜在的创始动物 在断奶时提供给研究人员进行DNA分析 Southern blotting用于基因分型。产生的ES细胞嵌合体 使用瑞士韦氏菌株作为宿主或通过 将ES细胞注射入C57BL/6囊胚。嵌合体被提供给 研究调查员在断奶后进行试验育种以确定 突变的生殖系传播。两者之间的聚合嵌合体 还可以通过TgESC设施制造小鼠品系。最近 提供的服务涉及制造嵌合体的新技术 四倍体桑椹产生100%来自ES细胞的胚胎。至 使研究人员能够在SPF动物体内安置无病原体的动物 在Skirball的宿舍，也提供了老鼠品系的重新派生。为 分离基因靶向的ES细胞，为用户提供DNA资源 与设施协商后准备的靶向构建物和 确认的基因分型分析。构建物被电穿孔进入ES细胞 在设施中准备的克隆，并使用 基因分型试验。将阳性同源重组子转移到 将突变的等位基因传递给小鼠的嵌合体单位。在……里面 此外，该设施为研究人员提供了广泛的 与生产转基因和敲除胚胎干细胞有关的服务 以及嵌合小鼠，如供应超排卵子捐赠者， 假孕母鼠，养鼠业培训和咨询 转基因/胚胎干细胞方法和载体设计。

The transgenic Mouse/ES Cell Chimeras Facility provides four basic services: production of transgenic (Tg) mice by zygote injection, production of embryonic stem (ES) cell chimeric animals, rederivation of mouse strains and isolation of gene-targeted ES cells following homologous recombination. For production of transgenic mice, users supply DNA constructs suitable for pro-nuclear injection into fertilized mouse eggs. Transgenic mice are produced in outbred Swiss Webster mice, inbred FVB/N mice or mice of defined genotype. Potential founder animals are provided to the research investigator at weaning for DNA analysis by Southern blotting for genotyping. ES cell chimeras as produced by morulae aggregation using the Swiss Webster strain as a host or by injection of ES cells into C57BL/6 blastocysts. Chimeras are provided to the research investigator at weaning for test breeding to determine germline transmission of the mutation. Aggregation chimeras between two strains of mice can also be made through the TgESC facility. A recent service being offered involved the new technique of making chimeras with tetraploid morulae to produce embryos that are 100% ES cell-derived. To enable investigators to house pathogen-free animals in the SPF animal quarters in Skirball, rederivation of mouse strains is also offered. For isolation of gene-targeted ES cells, users of the resource supply DNA targeting constructs prepared in consultation with the facility and a confirmed genotyping assay. Constructs are electroporated into ES cells prepared in the facility, and isolated clones are scored using the genotyping assay. Positive homologous recombinants are transferred to the chimera unit for transmission of the mutant allele into mice. In addition, the facility provides researchers with a wide array of services related to the production of transgenic and knockout ES cells and chimeric mice such as supply of superovulated egg donors, pseudopregnant females, training in mouse husbandry and consultation in transgenic/ES cell approaches and vector design.