Vector, Transgenic, and Imaging Core

载体、转基因和成像核心

• 批准号: 8794381
• 负责人: Viktor Kharazia
• 金额: $ 2.8万
• 依托单位: UNIVERSITY OF CALIFORNIA, SAN FRANCISCO
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/8794381
• 关键词: Alcohol consumption; Alcohols; Brain region; Cells; Complementary DNA; Dendrites; Detection; Drug Targeting; Equipment; Feedback; Gene Components; Gene Silencing; Gene Targeting; Genes; Genetic; Health Services Research; Histology; Housing; Human Resources; Image; In Situ Hybridization; In Vitro; Infection; Injection of therapeutic agent; Messenger RNA; Methods; Mission; Operative Surgical Procedures; Phosphorylation; Pilot Projects; Procedures; Proteins; RNA Interference; Relapse; Research; Research Personnel; Research Project Grants; Resolution; Services; Signaling Protein; Students; Techniques; Testing; Training; Transgenic Organisms; Viral; Viral Vector; Virus; Work; alcohol use disorder; binge drinking; cost; design; digital; image processing; immunocytochemistry; in vivo; interest; laser capture microdissection; novel; overexpression; protein expression; research study; response; screening; skills; small hairpin RNA; tool; vector

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.

矢量和成像核心（核心组件3）将提供集中的服务，供应和共享 所有建议使用病毒载体进行基因治疗的研究部门和试点项目的设备 通过RNA干扰在选择的大脑区域进行沉默。核心人员将设计有效的shRNA 分子，生产高滴度病毒，并进行体外和体内测试。核心人员还将 产生用于过表达目的基因的病毒。矢量和成像核心将继续 为研究组成部分和试点项目提供一系列组织学服务， 关于病毒注射的立体定位坐标、感染体积和感染细胞类型的反馈。的 Core将通过激光捕获显微切割结合定量分析来分析靶基因的表达。 PCR mRNA分析和免疫细胞化学。核心人员将继续进行实验， 信号蛋白的细胞定位，并将进一步扩展检测能力，通过执行高 树突中信号蛋白的分辨原位杂交和免疫细胞化学。额外的核心 职责包括：数字立体定位设备，成像设备的监督;立体定位培训 外科技术，组织学，成像和图像处理;指导学生和博士后 培训人员;以及为核心小组使用的技术制定标准程序。通过房屋矢量和 将成像服务集中在一个核心中，由于核心人员可以集中他们的技能， 这些技术是劳动密集型的，并且对几个研究项目通用。这将使调查人员 研究组成部分和试点项目，侧重于研究具体 候选蛋白质，同时避免技术错误和应用方法的不一致性， Vector和Imaging Core可以操纵这些蛋白质的表达。