Cell type differences in activity of the Streptomyces bacteriophage {phi}C31 integrase
2008
Maucksch, Christof | Aneja, Manish Kumar | Hennen, Elisabeth | Bohla, Alexander | Hoffmann, Florian | Elfinger, Markus | Rosenecker, Joseph | Rudolph, Carsten
Genomic integration by the Streptomyces bacteriophage {phi}C31 integrase is a promising tool for non-viral gene therapy of various genetic disorders. We investigated the {phi}C31 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 {phi}C31 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 stem cells, human lung, liver and cervix carcinoma cell lines. Up to 100-fold higher levels of recombination product was found in {phi}C31 integrase transfected A549 lung than Jurkat T cells. When the {phi}C31 integrase activity was normalized to the intracellular integrase mRNA levels, a 16-fold difference was found. As one possible inhibitor of the {phi}C31 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 {phi}C31 integrase activity.
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