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.
This Core has been a very valuable Component of the ACTG, allowing investigators to manipulate specific proteins in specific brain regions with genetic tools provided by the Core. The work of this Core is essential to the ACTG mission of understanding mechanisms by which novel proteins, identified as possible drug targets for treating alcohol use disorders, function in regulting binge drinking and relapse to alcohol use.
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