CORE--TRANSGENIC AND ANIMAL RESOURCE

描述：（直接摘自申请书）为了促进与PKD中心相关项目的全动物实验方法，现有的凯斯西部储备小鼠突变实验室将扩大，以满足中心研究人员的需求。胚胎干细胞培养和囊胚/原核注射技术是劳动密集型的，需要专门的培训和昂贵的设备。核心实验室将集中这些职责，从而使其更具生产力和成本效益。此外，核心将重新衍生，冷冻保存和管理各种项目所需的突变菌落的繁殖。该设施将负责：1。引进品系的再分化与低温保存集中育种、维护和储存所有由该设施制造或从其他来源的突变菌落进口的突变系，以及生产双突变体3。原核注射产生转基因小鼠二倍体生殖系能态胚胎干细胞的鉴定与维持电穿孔和筛选鉴定目标无性系靶系核型分析胚泡注射产生生殖系嵌合体与主要研究人员就育种方案、结构设计和实验策略进行磋商

Description: (Taken directly from the application) To facilitate whole animal experimental approaches for projects associated with the PKD Center, the existing Case Western Reserve Mouse Mutation Laboratory will be expanded to accommodate the needs of Center investigators. Embryonic stem (ES) cell culture and blastocyst/pronuclear injection techniques are labor intensive, requiring specialized training and expensive equipment. The Core laboratory will centralize these duties, thereby making it more productive and cost efficient. In addition, the Core will re-derive, cryopreserve and manage the breeding of mutant colonies needed for the various projects. The facility will be responsible for: 1. Re-derivation and cryopreservation of imported lines 2. Centralized breeding, maintenance and storage of all mutant lines either made by the facility or imported from other sources of mutant colonies as well as production of double mutants 3. Pronuclear injections to produce transgenic mice 4. Characterization and maintenance of diploid, germ line competent ES cells 5. Electroporation and selection to identify targeted clonal lines 6. Karyotype analysis of targeted lines 7. Blastocyst injections to produce germ line chimeras 8. Consultation with Principal Investigators regarding breeding schemes, construct design and experimental strategy