Abstract:
Genomic integration by the Streptomyces bacteriophage ΦC31 integrase is a promising tool for non-viral gene therapy of various genetic disorders. We investigated the ΦC31 integrase recombination activity in T cell derived cell lines, primary T lymphocytes and CD34+ haematopoietic stem cells in comparison to mesenchymal stem cells and cell lines derived from lung-, liver- and cervix-tissue. In T cell lines, enhanced long-term expression above control was observed only with high amounts of integrase mRNA. Transfections of rC31 integrase plasmids were not capable of mediating enhanced long-term transgene expression in T cell lines. In contrast, moderate to high efficiency could be detected in human mesenchymal stemcells, human lung, liver and cervix carcinoma cell lines. Up to 100-fold higher levels of recombination product was found in ΦC31 integrase transfected A549 lung than Jurkat T cells. When the rC31 integrase activity was normalized to the intracellular integrase mRNA levels, a 16-fold difference was found. As one possible inhibitor of the ΦC31 integrase, we found 3- to 5-fold higher DAXX levels in Jurkat than in A549 cells, which could in addition to other yet unknown factors explain the observed discrepancy of ΦC31 integrase activity.
