CORE--MAMDC GENE TARGETING CORE FACILITY

核心--MAMDC基因靶向核心设施

• 批准号: 6100351
• 负责人: Carl A. Pinkert
• 金额: $ 13.38万
• 依托单位: UNIVERSITY OF ALABAMA AT BIRMINGHAM
• 依托单位国家: 美国
• 财政年份: 1999
• 资助国家: 美国
• 起止时间: 1999-01-01 至 1999-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6100351
• 关键词: arthritis; biomedical facility; biotechnology; cell bank /registry; cell line; embryonic stem cell; gene targeting; genetically modified animals; laboratory mouse; microinjections; model design /development; musculoskeletal disorder; transfection

The proposed facility in this UAB Multipurpose Arthritis and Musculoskeletal disease Center (MAMDC) application provides for a valuable and dynamic set of resources reflecting the intensity and direction of research endeavors and efforts at The University of Alabama at Birmingham (UAB). The UAB Transgenic Animal/ES Cell Resource (Resource) provides state-of-the-art gene transfer technology including the production of both gain-of-function and/or loss-of-function ("knockout") mouse models. The Resource produces transgenic mice using both DNA microinjection and embryonic stem cell (ES cell) transfer methodologies. However, the costs for manipulation and transfer of embryonic stem cells is considerable greater than that for DNA microinjection, hence, the absolute costs preclude the utilization of this valuable modeling system. Currently, investigators interested in ES cell technology must establish in vitro ES cell techniques within their laboratory. Once cell lines are propagated, the PI is responsible not only for all preliminary in vitro testing but for the costs associated with in vivo testing of cell lines. Thus, the inherent initial costs effectively limit novel experimentation. The proposed core will provide a more comprehensive program for the development of ES cell-derived mouse models which will be useful to explore in vivo modulation of human gene expression and as a preclinical experimental model system. Therefore, the specific aim of this proposal is to reduce the developmental costs associated with ES cell culture and manipulation, leading to the production of pathogen-free mouse models for MAMDC investigators. This proposal will be used specifically to support: a) setup costs for a full-time ES cell culture program, b) cell transfection and culture, c) clone preparation for investigators, d) propagation of investigator identified cell clones for founder stem cell-derived or "knockout" mouse production, e) a repository for ES cell lines and investigator clones, f) evaluation and implementation of new technologies as they become relevant over the duration of the proposed program, and g) maintenance and testing of cell lines obtained from different investigators through to cell injection into mouse embryos, thereby characterizing the relative utility and efficiency associated with specific lines and cell passages.

UAB多功能关节炎和 肌肉骨骼疾病中心（MAMDC）的应用提供了 有价值而动态的资源集，反映了强度和 阿拉巴马大学的研究努力和努力方向 在伯明翰（UAB）。 UAB转基因动物/ES细胞资源 （资源）提供最先进的基因转移技术，包括 功能获得和/或功能丧失的产生 （“敲除”）鼠标模型。 资源使用 DNA显微注射和胚胎干细胞（ES细胞）转移 方法论。 但是，操纵和转移的成本 胚胎干细胞比DNA大得多 因此，绝对成本排除了使用 这个有价值的建模系统。 目前，调查人员对 ES细胞技术必须建立体外ES细胞技术 他们的实验室。 一旦细胞系传播，PI就负责 不仅是所有初步的体外测试，还为成本 与细胞系的体内测试有关。 因此，固有 初始成本有效地限制了新颖的实验。 提议 核心将为ES的发展提供更全面的计划 细胞来源的鼠标模型，可用于探索体内 人类基因表达和临床前实验的调节 模型系统。 因此，该提议的具体目的是减少 与ES细胞培养和 操纵，导致无病原体小鼠模型的产生 对于MAMDC调查人员。 该建议将专门用于 支持：a）全日制ES培养计划的设置成本，b）细胞 转染和培养，c）研究人员的克隆准备，d） 研究者的传播确定了创始人茎的细胞克隆 细胞来源或“淘汰”小鼠的产生，e）ES细胞的存储库 线和调查员克隆，f）评估和实施新的 技术在拟议的期限内变得相关 程序，g）从 不同的研究者通过注射小鼠胚胎， 从而表征相对效用和相关的效率 具有特定的线条和单元格。