CORE--MAMDC GENE TARGETING CORE FACILITY

该UAB多用途关节炎的拟议设施， 肌肉骨骼疾病中心（MAMDC）的应用程序提供了一个 一套宝贵而充满活力的资源，反映了 亚拉巴马大学的研究方向和努力 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 产生功能获得和/或功能丧失 （"敲除"）小鼠模型。 转基因小鼠的生产使用 DNA显微注射和胚胎干细胞（ES细胞）移植 方法论。 然而，操纵和转让的成本 胚胎干细胞比DNA干细胞 显微注射，因此，绝对成本排除利用 这个有价值的建模系统 目前，调查人员对 ES细胞技术必须建立在体外ES细胞技术内， 他们的实验室。 一旦细胞系繁殖，PI负责 不仅用于所有初步的体外试验， 与细胞系的体内测试相关。 因此， 初始成本有效地限制了新的实验。 拟议 core将为ES的开发提供更全面的方案 细胞衍生的小鼠模型，这将有助于在体内探索 调节人类基因表达，并作为临床前实验 模型系统 因此，这项建议的具体目的是减少 与ES细胞培养相关的开发成本， 操作，导致产生无病原体的小鼠模型 为MAMDC调查员。 该提案将专门用于 支持：a）全职ES细胞培养计划的设置成本，B）细胞 转染和培养，c）用于研究者的克隆制备，d） 研究者鉴定的用于创始干细胞的细胞克隆的繁殖 细胞衍生或"敲除"小鼠生产，e）ES细胞库 f）评价和实施新的 技术，因为它们在拟议的 程序，和g）维持和测试从 不同的研究人员通过将细胞注射到小鼠胚胎中， 从而表征相关的相对效用和效率 特定的细胞系和细胞传代。

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.