Design and construction of integrase deficient lentiviral vector for gene therapy

Gene therapy using gene targeting approach is one of the best therapeutic methods for treatment of monogenic disorders. In contrast to application of viral vectors, gene targeting has low efficiency. The scope of this study is to design and construct a lentiviral vector without integration into the genome, but entering into the cell nucleus with high efficiency. In this study after transduction of 293T cells with native virus and recombinant virus, eGFP percentage in transduced cells with native virus was decreased slightly two weeks after viral treatment. In contrast, in negative integrase viruses a dramatic decrease was observed. Results shows that recombinant virus genome remains as episomal and integrates with lower efficiency into the host genome which has the lowest undesired effect on expression of other genes of cell. Defective gene can be replaced by gene entered into the cell nucleus without any viral genome portion integration into the host genome.