486 Rapid Deployment Plasmid Production Process, Combining Inducible High Yield Fed Batch Fermentation Process with Novel Autolytic Plasmid DNA Purification 484 Safety of q>C31 Integrase Expression in[.]
484 Safety of q>C31 Integrase Expression in Mouse Liver Following Hydrodynamic Injection Christopher L Chavez, Lauren E Woodard , Annahita Keravala, Vanessa Gabrovsky 'Genetics, Stanford Stanford CA tpC3l intcgrasc, a site-specific integrase from the bacteriophage tpC31, catalyzes chromosomal integration of plasmid DNA at ' pseudo' attP sites in mammalian genomes tpC3l integrase has been used to achieve integration and long-term, robust expression of therapeutic genes in many tissues, including mouse liver No immune reaction or tumors have been observed in these studies We are undertaking additional studies to examine more thoroughly the safety of using tpC31 integrasc in gene therapy of the liver The predicted short duration of integrase expression would be a safety feature by limiting exposure of cells to the reeombinase In order to test this hypothesis, we are determining the longevity ofintegrase protein afler hydrodynamic injection of an integrase expression vector in the mouse liver The longevity of integrase expression will be determined by removing the liver at various time points, isolating total protein for Western blotting from half, and sectioning the remaining half for immunohistochemistry with an antibody detecting