Nucleic Acids Research Advance Access originally published online on August 21, 2008
Nucleic Acids Research 2008 36(17):5462-5471; doi:10.1093/nar/gkn532
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Nucleic Acids Research, 2008, Vol. 36, No. 17 5462-5471
© 2008 The Author(s)
This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/2.0/uk/) which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.
Nucleic Acid Enzymes |
Cell type differences in activity of the Streptomyces bacteriophage 
C31 integrase
1Department of Pediatrics, Ludwig-Maximilians-University, 80337 Munich and 2Free University of Berlin, Department of Pharmacy, 14166 Berlin, Germany
*To whom correspondence should be addressed. Tel: +49 89 5160 7711; Fax: +49 89 5160 7846; Email: carsten.rudolph{at}med.uni-muenchen.de
Received June 30, 2008. Revised July 31, 2008. Accepted August 1, 2008.
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.