(See inslructions): Core C is composed of two modules: the Vector Module and the Animal Module. This core will provide the resources and expertise needed for the efficient generation of a wide array of gene-transfer technologies, genotyping ofthe growing numbers of transgenic animal models (breeding colonies) used by Program Project Investigators, and for phenotypic characterization of animal models including telemetry-based blood pressure measurements, assessment of renal and vascular functions, and histology. The Vector Module will develop and construct HIV-1-based Lentivirus vectors capable of targeting specific cells using endothelial, smooth muscle, adipocytes and transport epithelial cell-specific promoters (e.g., SM22 alpha, VECAD, AP2, THP, NKCC2) to overexpress and suppress (Antisense and shRNA) genes of interest along with corresponding controls, i.e., GFP, catalytically inactive enzymes and non-specific shRNA. The Vector Module will isolate, titer and concentrate purified viruses as well as assist investigators in administering viral vectors, assessing efficiency of viral-mediated gene expression, and genotyping to ensure the quality of breeding colonies for Program Project Investigators. The Animal Module will provide technologies and services including measurement of arterial blood pressure by radiotelemetry and tail-cuff methodology, assessment of renal function (GFR), preparation of tissues for histology/pathology and immunohistochemistry, and assistance in measuring vascular reactivity (wire and pressure myographs). An understanding ofthe molecular mechanism by which CYP-derived eicosanoids contribute to the regulation of blood pressure is contingent upon the identification of appropriate methods of gene manipulation and ability to subsequently assess pathophysiological changes. The centralization of these services within Core C will provide efficient, standardized and accurate utilization of molecular and physiological resources and procedures that are essential for each of the projects.
The ability to specifically target gene expression in a cell-specific manner is essential to discerning interactions between components of blood pressure control, including the kidney, the vasculature and adipocytes. Likewise it is vital to be able to accurately measure pathophysiological changes in response to genetic, molecular, and pharmacological interventions.
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