CORE--GENE TRANSFER

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The function of the Gene Transfer Core is to provide a facility for the construction, screening, large-scale purification, and titrating of replication-defective, recombinant adenoviruses. Recombinant adenoviruses provide a reliable method for high efficiency gene transfer into adult cardiocytes and for directly altering the phenotype. Specifically, the core will 1) maintain lines of human 293 endothelial kidney cells [HEK], 2) amplify and maintain purified stocks of adenovirus shuttle plasmids and adenovirus backbone plasmids, 3) transfect recombinant adenoviral DNA into 293 HEK cells, 4) propagate recombinant adenoviruses in 293 HEK cells, 5) plaque-purify recombinant adenoviruses and perform large-scale purifications, and 6) titer purified adenoviruses by plaque assays in 293 HEK cells. The core will also be responsible for long-term storage of recombinant adenoviruses and will assist individual investigators in carrying out gene transfer protocols in adult cardiocytes. A full-time research specialist has been trained in all of the procedures and techniques involved in running this core, and will be supervised by the core director.