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- Title
5-Azacytidine prevents transgene methylation in vivo.
- Authors
Ianni, M Di; Terenzi, A; Perruccio, K; Ciurnelli, R; Lucheroni, F; Benedetti, R; Martelli, M F; Tabilio, A
- Abstract
Retroviral sequence can silence transgene expression in vitro and in vivo. We report that this effect can be efficiently prevented by in vivo administration of the demethylating agent 5-azacytidine (aza-C). We engineered the U937 human cell line with a retroviral vector consisting of the thymidine kinase suicide gene (tk), which induces sensitivity to ganciclovir (gcv) and through an IRES sequence, the bacterial beta-galactosidase gene (lacZ) as a marker gene. About 90% of the U937 cells expressed the transgene. By injecting the transduced U937 cells in severe combined immunodeficient disease (SCID) mice, we generated a tumour which, during in vivo treatment with aza- C, maintained the high expression of lacZ and tk genes at the baseline values. LacZ-positive cells in the tumour masses after death was weak (1–2%) in the control group, while in mice treated with aza-C it was maintained at 90%. The delay in tumour onset was significanly longer when animals were treated with both aza-C and gcv (P < 0.0001) compared with animals treated with gcv or with aza-c alone. the prevention of silencing phenomena has important implications for gene therapy, because an efficient transduction associated with appropriate drug therapy, might be a powerful strategy for successful application of gene therapy protocols.
- Subjects
TRANSGENE expression; DNA; METHYLATION; RETROVIRUSES; GENETIC vectors
- Publication
Gene Therapy, 1999, Vol 6, Issue 4, p703
- ISSN
0969-7128
- Publication type
Article
- DOI
10.1038/sj.gt.3300848