A strategy for enhancing the transcriptional activity of weak cell type-specific promoters

Citation
Dm. Nettelbeck et al., A strategy for enhancing the transcriptional activity of weak cell type-specific promoters, GENE THER, 5(12), 1998, pp. 1656-1664
Citations number
42
Categorie Soggetti
Molecular Biology & Genetics
Journal title
GENE THERAPY
ISSN journal
09697128 → ACNP
Volume
5
Issue
12
Year of publication
1998
Pages
1656 - 1664
Database
ISI
SICI code
0969-7128(199812)5:12<1656:ASFETT>2.0.ZU;2-2
Abstract
Cell type- and tissue-specific promoters play an important role in the deve lopment of site-selective vectors for gene therapy. A large number of highl y specific promoters has been described, but their applicability is often h ampered by their inefficient transcriptional activity. In this study, we de scribe a new strategy for enhancing the activity of weak promoters without loss of specificity. The basic principle of this strategy is to establish a positive feedback loop which is initiated by transcription from a cell typ e-specific promoter. This was achieved by using a cell type-specific promot er to drive the simultaneous expression of the desired effector/reporter ge ne product and a strong artificial transcriptional activator which stimulat es transcription through appropriate binding sites in the promoter. Using a VP16-LexA chimeric transcription factor, we show that this approach leads to a 14- to > 100-fold enhancement of both and the gastrointestinal-specifi c sucrase-isomaltase promoter while maintaining approximately 30- to > 100- fold cell type specificity.