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Nucleic Acids Research, 2000, Vol. 28, No. 5 1120-1125
© 2000 Oxford University Press

The tetracycline-responsive promoter contains functional interferon-inducible response elements

Andreas Rang and Hans Will*

Heinrich-Pette-Institut, Martinistraße 52, D-20251 Hamburg, Germany

Tetracycline (tet)-responsive expression vectors allow controlled inducible expression of proteins in mammalian cells. This system is widely used for experimental research both in vivo and in vitro. In our attempts to use this system to study the antiviral effect of IFN{alpha} on hepatitis B virus, we discovered an unexpected feature of the tet-responsive promoter (tet promoter) of the currently available expression vectors. IFN{alpha} was found to stimulate tet promoter activity after transient transfection in a dose- and cell type-dependent manner. By sequence inspection, an IFN{alpha}-stimulated response element (ISRE)-like sequence was identified in the linker regions located between the heptameric tet operator sequences. Gel shift assays revealed binding of IFN-stimulated gene factors to these sequences, indicating that they mediate the IFN{alpha}-mediated promoter stimulation. These data demonstrate an unexpected feature of the tet-responsive expression system which needs to be taken into acount when using this system for analysis of cytokine functions in vitro and in vivo. The data also imply that the tet promoter-based expression system can be rendered non-responsive to IFN{alpha} by mutagenesis of the ISREs and this may be essential when considering gene therapy in vivo.

* To whom correspondence should be addressed. Tel: +49 40 48051 221; Fax: +49 40 48051 222; Email: will@hpi.uni-hamburg.de


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