Core--Inhalation /transgenic mouse

核心--吸入/转基因小鼠

• 批准号: 7056718
• 负责人: DAVID R. HEMENWAY
• 金额: $ 26.29万
• 依托单位: UNIVERSITY OF VERMONT & ST AGRIC COLLEGE
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/7056718
• 关键词: animal colony; biomedical facility; genetically modified animals; laboratory mouse; pulmonary fibrosis /granuloma

The Specific Aims of the Inhalation Core are to provide housing of mice prior to inhalation experiments in a pathogen-free environment and controlled exposures to asbestos and ovalbumin (OVA). The objective of the Transgenic Mouse Core is to provide transgenic mice to the members of the Program Project. The generation, maintenance and screening of new and previously generated conventional transgenic mice will be centralized in this Core. In addition, this Core aims to develop a new experimental approach that involves stable integration of bacterial artificial chromosomes (BACs) in the mouse genome. The Specific Aims of this Core are: i. To provide a Core service for the generation of new transgenic mice (F2 C3H x C57Bl/6) expressing specific transgenes in pulmonary epithelial cells. The new transgenic mice that are proposed in this Program Project will be generated in the University of Vermont Transgenic Mouse Core, directed by Dr. Rincon. ii. To centralize the backcross process required to obtain each of the new and previously generated transgenic mice in two specific strain backgrounds. The new F2 C3H x C57Bl/6 transgenic mice will be backcrossed for seven generations with wild type inbred mice (C57Bl/6 and Balb/c) to obtain homogeneous genetic background. iii. To screen transgenic mice by molecular analyses of genomic DNA (PCR, slot blot) and provide the necessary number of mice to the investigators of the Program. The screening process will also be centralized to prevent variability and increase efficiency and reproducibility of the results from different projects. iv. To develop new applications involving the use of transgenic mice to study lung fibrosis. An alternative approach to complement the classical transgenic mouse models will be developed during the Program funding period: the generation of transgenic mice by BACs.

吸入器核心的具体目标是在无病原体的环境中进行吸入实验之前为小鼠提供住所，并控制石棉和卵清蛋白(OVA)的暴露。转基因小鼠核心的目标是为项目成员提供转基因小鼠。新的和以前产生的常规转基因小鼠的产生、维护和筛选将集中在该中心。此外，该核心旨在开发一种新的实验方法，涉及将细菌人工染色体(BAC)稳定整合到小鼠基因组中。该中心的具体目标是：1.为在肺上皮细胞中表达特定转基因的新转基因小鼠(F2C3Hx C57BL/6)的产生提供核心服务。该计划项目中提出的新转基因小鼠将在佛蒙特大学转基因小鼠核心中产生，由林肯博士指导。二、集中回交过程，以获得两个特定品系背景中的每一只新的和先前产生的转基因小鼠。将新的F2C3Hx C57BL/6转基因小鼠与野生型近交系小鼠(C57BL/6和Balb/c)回交7代，以获得纯合的遗传背景。三、通过基因组DNA的分子分析(聚合酶链式反应，狭缝杂交)筛选转基因小鼠，并为该计划的研究人员提供必要的小鼠数量。筛选过程也将集中进行，以防止多变，提高不同项目结果的效率和可重复性。四、开发新的应用，包括使用转基因小鼠来研究肺纤维化。在计划资助期内，将开发一种替代方法来补充经典的转基因小鼠模型：由BAC产生转基因小鼠。