title: Cell type differences in activity of the Streptomyces bacteriophage phiC31 integrase.
abstract: Genomic integration by the Streptomyces bacteriophage C31 integrase is a promising tool for non-viral gene therapy of various genetic disorders. We investigated the 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 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 C31 integrase transfected A549 lung than Jurkat T cells. When the C31 integrase activity was normalized to the intracellular integrase mRNA levels, a 16-fold difference was found. As one possible inhibitor of the 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 C31 integrase activity.
authors: Department of Pediatrics, Ludwig-Maximilians-University, 80337 Munich, Germany.
authors: Christof
authors: C
authors: Maucksch
authors: Manish Kumar
authors: MK
authors: Aneja
authors: Elisabeth
authors: E
authors: Hennen
authors: Alexander
authors: A
authors: Bohla
authors: Florian
authors: F
authors: Hoffmann
authors: Markus
authors: M
authors: Elfinger
authors: Joseph
authors: J
authors: Rosenecker
authors: Carsten
authors: C
authors: Rudolph
mesh_terms: D048868:Adaptor Proteins, Signal Transducing / Q000032:analysis; D001435:Bacteriophages / Q000201:enzymology*; D002460:Cell Line; D045744:Cell Line, Tumor; D002478:Cells, Cultured; D056970:Co-Repressor Proteins; D015870:Gene Expression; D017930:Genes, Reporter; D006801:Humans; D019426:Integrases / Q000378:metabolism*; D019169:Jurkat Cells; D018832:Molecular Chaperones; D009687:Nuclear Proteins / Q000032:analysis; D010957:Plasmids / Q000235:genetics; D016133:Polymerase Chain Reaction; D011995:Recombination, Genetic; D013302:Streptomyces / Q000821:virology; D013601:T-Lymphocytes / Q000378:metabolism; D014162:Transfection
chemical_list: D048868:Adaptor Proteins, Signal Transducing; D056970:Co-Repressor Proteins; C106985:DAXX protein, human; D018832:Molecular Chaperones; D009687:Nuclear Proteins; D019426:Integrases