Vector, Transgenic, and Imaging Core

向量和成像核心(核心构成部分3)将提供集中服务、用品和共享 向所有建议使用基因病毒载体的研究部门和试点项目提供设备 在选定的大脑区域通过RNA干扰沉默。核心人员将设计有效的shRNA 分子，产生高滴度病毒，并进行体外和体内测试。核心人员也将 产生病毒以过度表达感兴趣的基因。向量和成像核心将继续 为研究部分和试点项目提供一系列组织学服务，方法是 关于病毒注射、感染量和感染细胞类型的立体定位坐标的反馈。这个 CORE将通过激光捕获显微切割结合定量分析目标基因的表达 聚合酶链式反应基因分析及免疫细胞化学方法。核心人员将继续在 信号蛋白的细胞定位，并将通过执行高性能进一步扩展检测能力 树突状细胞信号蛋白的分辨原位杂交和免疫细胞化学。附加核心 职责包括：监督数字立体定位仪设备、成像设备；培训立体定位仪 外科技术、组织学、成像和图像处理；指导学生和博士后 受训人员；为核心使用的技术制定标准程序。通过住房向量和 影像服务在一个核心，效率和成本可以优化，因为核心人员可以专注于他们的技能 关于劳动密集型和对几个研究项目普遍适用的技术。这将使调查人员能够 研究组成部分和试点项目，侧重于具体研究中涉及的科学问题 候选蛋白质，同时避免在应用方法时出现技术错误和不一致 载体和成像核心可以操纵这些蛋白质的表达。

The Vector and Imaging Core (Core Component 3) will provide centralized services, supplies, and shared equipment to all Research Components and Pilot Projects that propose use of viral vectors for gene silencing by RNA interference in select brain regions. Core personnel will design effective shRNA molecules, produce high titer viruses, and perform in vitro and in vivo testing. Core personnel will also produce viruses for overexpression of genes of interest. The Vector and Imaging Core will continue to provide a range of histological services for the Research Components and Pilot Projects, by providing feedback on stereotaxic coordinates of viral injections, volume of infections, and types of infected cells. The Core will analyze expression of targeted genes by laser capture microdissection combined with quantitative PCR mRNA analysis, and by immunocytochemistry. Core personnel will continue to perform experiments on the cellular localization of signaling proteins, and will further extend detection capabilities by performing high resolution in situ hybridization and immunocytochemistry for signaling proteins in dendrites. Additional Core responsibilities include: oversight of digital stereotaxic equipment, imaging equipment; training in stereotaxic surgical techniques, histology, imaging and image processing; supervising students and postdoctoral trainees; and generating standard procedures for techniques used by the Core. By housing vector and imaging services in one core, efficiency and cost can be optimized since core personnel can focus their skills on techniques that are labor intensive and universal to several research projects. This will allow investigators of Research Components and Pilot Projects to focus on scientific questions involved in studies of specific candidate proteins, while avoiding technical errors and inconsistency in application of methods that the Vector and Imaging Core can perform to manipulate expression of these proteins.