TRANSGENIC MOUSE CORE

转基因小鼠核心的目标是为每个项目的研究人员提供帮助 转基因小鼠的设计和生产。为了实现这一目标，核心将执行以下操作 功能： 具体目标1.设计和构建转基因并开发基因分型方法。核心人员将 就每个拟议的转基因模型的目标咨询项目负责人。适当的DNA 然后将使用最佳启动子和编码序列来设计构建体。所需的基因组或 将获得目的基因的cDNA克隆，并从所需启动子的下游进行克隆。 转基因DNA将被提纯用于显微注射，并将设计和测试基因分型分析。 具体目的2.通过显微注射转基因DNA获得转基因创建者。受精卵来自 将C57BL/6系小鼠用于转基因构建物DNA的原核注射。潜在地 转基因后代将通过针对特定目标开发的基因分型分析进行鉴定1.创始小鼠 将以将转基因传播给后代的能力为特征。独立的数量 将确定每个创建者的染色体整合位置。具有生殖系能力的小鼠，每个小鼠都有一个 整合在单个基因组位置的转基因将被交付给项目负责人。 具体目标3.协助确定转基因小鼠的特征。定量逆转录酶 聚合酶链式反应(RT-PCR)分析将被设计用于特异性地检测转基因mRNAs和 然后用来确定转基因基因表达的组织分布。如果转基因表达是 低层核心人员将协助项目负责人找出低表达和优化的原因 转基因设计以克服这一问题。 转基因小鼠越来越多地被用于解决衰老和骨骼研究中的问题。实验在 转基因小鼠使研究人员能够确定在细胞系和原代细胞中观察到的现象 培养也会在体内发生。此外，转基因小鼠可以用来检验那些不能 在任何体外系统中都能令人信服地解决。因此，及时高效地生产转基因小鼠是 对该计划的总体目标至关重要，该目标是提高对糖尿病的病理生理学的理解 治疗骨质疏松症的骨脆性综合征，从而使其治疗合理化和最优化。

The goal of the Transgenic Mouse Core is to provide the investigators of each project with assistance in the design and production of genetically modified mice. To achieve this goal, the core will perform the following functions: Specific Aim 1. Design and construct transgenes and develop genotyping assays. Core personnel will consult with the project leaders regarding the goal for each proposed transgenic model. Appropriate DNA constructs will then be designed using optimal promoter and coding sequences. The required genomic or cDNA clones for the gene of interest will be acquired and cloned downstream from the desired promoter. Transgene DNA will be purified for microinjection, and genotyping assays will be designed and tested. Specific Aim 2. Generate transgenic founders via microinjection of transgene DNA. Fertilized eggs from the C57BL/6 strain of mice will be used for pro-nuclear injection of transgene construct DNA. Potentially transgenic offspring will be identified using genotyping assays developed in Specific Aim 1. Founder mice will be characterized for the ability to transmit the transgene to offspring. The number of independent chromosomal integration sites in each founder will be determined. Germline-competent mice, each with a transgene integrated at a single genomic locus, will be delivered to project leaders. Specific Aim 3. Assist in the characterization of transgenic mice. Quantitative reverse-transcriptase polymerase chain reaction (RT-PCR) assays will be designed to specifically detect transgene mRNAs and then used to determine the tissue distribution of transgene mRNA expression. If transgene expression is low, core personnel will assist the Project Leader in identifying reasons for low expression and optimizing transgene design to overcome the problem. Transgenic mice are increasingly used to address questions in aging and skeletal research. Experiments in transgenic mice allow investigators to determine whether phenomena observed in cell lines and primary cell cultures also occur in vivo. In addition, transgenic mice can be used to test hypotheses that cannot be convincingly addressed in any in vitro system. Thus timely and efficient production of transgenic mice is essential to the overall goal of the Program which is to improve the understanding of the pathophysiology of the bone fragility syndrome of osteoporosis and thereby rationalize and optimize its treatment.