Transgenic Mouse Core

本计划项目资助申请的总体目标是阐明自身免疫的机制。 心脏和肺移植中的V型胶原。使用动物模型来研究发病机制是必要的 加深了我们对这一重要临床问题的认识。为此，转基因技术的研究重点 MICE CORE(CORE B)是为转基因小鼠的世代提供技术专长 品系，并保持这些独特的小鼠品系的群体，以便为 针对所提议的三个项目的目标的实验。为此，核心B的第一部分将 利用威斯康星大学生物技术中心(UWBC)转基因动物设施的人员 (Taf)至(1)在胚胎干细胞中进行基因打靶实验，以产生ES细胞克隆 转基因等位基因和囊胚显微注射产生新的小鼠突变株 这些突变的等位基因，(2)重新衍生从其他机构或 目前被安置在非SPF动物设施中，以便该项目的老鼠种群可以被安置在 威斯康星大学新开放的无特定病原体的组织，以及(3)长期冷冻保存 定期保存和安全保存本提案中使用的所有转基因小鼠品系。在第二个 作为核心B的一部分，核心工作人员将(1)核实所有为 项目和某些菌株的可诱导组织特异性表达，(2)维持所有 转基因小鼠品系，从而可以提供所需数量的小鼠种群 根据个别项目的实验需要，将其添加到各个项目。此核心将为以下人员提供特定服务 本提案中的三个项目中的每一个。这一核心提供的服务将是实现 这项拨款申请的总体目标。

The overall goal of this Program Project grant application is to elucidate the mechanisms of autoimmunity to type V collagen in heart and lung transplants. The use of animal models to study mechanism is essential for advancing our understanding of this important clinical problem. To this end, the focus of the Transgenic Mouse Core (Core B) is to provide the technical expertise for the generation of genetically modified mouse strains and to maintain a colony of these unique mouse strains such that mice are generated for the experiments that address the aims of the proposed three Projects. To do so, the first part of Core B will utilize personnel in the University of Wisconsin Biotechnology Center's (UWBC) Transgenic Animal Facility (TAF) to (1) carry out gene targeting experiments in embryonic stem cells to produce ES cell clones with genetically altered alleles and the blastocyst microinjections to generate new mouse mutant strains carrying these mutant alleles, (2) to rederive genetically modified strains that are obtained from other institutions or are housed presently in non-SPF animal facilities so that the mouse stocks for this project can be housed in a newly opened specific pathogen free vivarium at the University of Wisconsin, and (3) cryopreserve for long term storage and safe keeping all the genetically modified mouse strains used in this proposal. In the second part of Core B, Core staff will (1) verify the genetic identity of all the mutant mouse strains rederived for the projects and the inducible tissue specific expression of certain strains, (2) maintain a breeding colony of all the genetically modified mouse strains such that the required numbers of the mouse stocks can be supplied to the individual Projects as needed for their experimentation. This Core will provide specific services for each of the three projects in this proposal. The services provided by this Core will be essential for achieving the overall goals of this grant application.