CORE--MAMDC GENE TARGETING CORE FACILITY

在这个多用途关节炎和UAB的拟议设施 肌肉骨骼疾病中心(MAMDC)应用程序提供 有价值的和动态的资源集合，反映了强度和 阿拉巴马大学的研究方向和努力方向 在伯明翰(UAB)。UAB转基因动物/胚胎细胞资源 (资源)提供最先进的基因转移技术，包括 功能增益和/或功能损失的产生 (“基因敲除”)小鼠模型。该资源生产转基因小鼠使用 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要大得多。 因此，微量注射的绝对成本使其无法利用。 这个有价值的建模系统。目前，有兴趣的调查人员 胚胎干细胞技术必须建立在体外胚胎干细胞技术 他们的实验室。一旦细胞系被繁殖，PI就负责 不仅是所有初步的体外试验，而且还有成本 与体内细胞系测试有关。因此，固有的 初始成本有效地限制了新颖的实验。建议数 CORE将为ES的发展提供更全面的方案 用于活体探索的细胞源小鼠模型 人类基因表达的调控及作为临床前实验 模型系统。因此，这项提议的具体目的是减少 与胚胎干细胞培养和发育相关的成本 操纵，导致产生无病原体的小鼠模型 给MAMDC的调查人员。这项提案将专门用于 支持：a)全日制ES细胞培养计划的设置费用，b)细胞 转染法和培养法，c)为研究人员准备克隆，d) 研究人员鉴定的方正干细胞克隆的繁殖 细胞衍生或“敲除”小鼠的生产，e)胚胎干细胞的储存库 线路和调查员克隆，f)评价和实施新的 技术，因为它们在提议的 编程，以及g)维护和测试从 从不同的研究人员到将细胞注射到小鼠胚胎， 从而表征相关联的相对效用和效率 有特定的线条和细胞通道。

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.