CORE--TRANSGENIC ANIMAL FACILITY

该计划中的几个项目将利用以下组合： 转基因和基因靶向小鼠研究信号机制 导致心脏肥大和/或扩张型心肌病。 在 在某些情况下，转基因方法将用于确定的作用， 一种激活不同分子的候选信号分子， 肥大的形态学或生理学特征。 心脏- 特异性表达将通过定义明确的启动子实现， 在转基因小鼠中驱动心脏限制性表达。 过表达 用于拯救研究的野生型蛋白质的表达（MLP），或 将使用主要活性蛋白质。 或者，基因- 靶向策略（通过ES细胞中的同源重组）是 目前在我们的项目中全面实施。 钱氏实验室拥有广泛的 在生产、鉴定、育种和 转基因和基因靶向小鼠模型的表征 心脏生理学和疾病。 该核心股将提供服务，协助设计 适当的构建体，在培养的细胞中测试构建体， 模型，以及随后的基因靶向和转基因 携带感兴趣的转基因的小鼠。 最后，创始人老鼠 将在核心动物设施中进行识别、繁殖和饲养。 在 在某些情况下，转基因系将用于培养 将与细胞一起制备的心肌细胞 生物学核心。 因此，该核心单位的目标有五个方面： 1)在体外系统中设计和测试适当的转基因; 2)产生合适的转基因和基因靶向小鼠品系 项目中的各种项目; 3)确定创始人并培育/维持适当的品系， 小鼠; 4)通过同源重组敲除特定的候选调控基因， 重组 5)为了繁殖和产生心脏限制所需的小鼠品系， 或通过CRE-lox策略的条件基因靶向。

Several of the projects in the Program will utilize a combination of transgenic and gene-targeted mice to study the signaling mechanisms which lead to cardiac hypertrophy and/or dilated cardiomyopathy. In certain cases, transgenic approaches will be used to identify the role of a candidate signaling molecule in the activation of distinct molecular, morphological, or physiological features of hypertrophy. Cardiac- specific expression will be achieved via well-defined promoters that can drive cardiac-restricted expression in transgenic mice. Over-expression of wild type proteins for rescue studies (MLP), or expression of dominantly active proteins will be utilized. Alternatively, gene- targeting strategies (via homologous recombination in ES cells) are currently fully operative in our Program. The Chien lab has extensive experience in the generation, identification, breeding, and characterization of both transgenic and gene-targeted mouse models of cardiac physiology and disease. This Core Unit will provide services to assist in the design of the appropriate constructs, the testing of the constructs in cultured cell models, and the subsequent generation of gene-targeted and transgenic mice which harbor transgenes of interest. Finally, the founder mice will be identified, bred, and maintained in a core animal facility. In certain cases, the transgenic lines will be utilized for cultured myocardial cells which will be prepared in conjunction with the Cell Biology core. Accordingly, the objectives of the core unit are five-fold: 1) To design and test appropriate transgenes in in vitro systems; 2) To generate appropriate lines of transgenic and gene-targeted mice for various projects in the Program; 3) To identify founders and to breed/maintain appropriate lines of mice; 4) To knockout specific candidate regulatory genes via homologous recombination. 5) To breed and generate lines of mice required for cardiac restricted or conditional gene targeting via CRE-lox strategies.