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Nucleic Acids Research, 2003, Vol. 31, No. 23 e152
© 2003 Oxford University Press

Article

Suppression of gene expression by a cell-permeable Tet repressor

Alison Mortlock, Walter Low and Andrea Crisanti*

Biogeny PLC and Department of Biology and Biochemistry, SAF Building, Imperial College, London SW7 2AZ, UK

*To whom correspondence should be addressed. Tel: +44 207 594 5412; Fax: +44 207 594 5439; Email: a.drcrisanti{at}imperial.ac.uk

Engineered transcription factors designed to selectively activate or repress endogenous genes have great potential in medical and biotechnological applications. Ultimately, their success will depend on the development of efficient delivery systems. We show here that a chimeric tetracycline- controlled transcription factor, encompassing the Tet repressor (TetR) from the tetracycline-resistance operon (tet from Escherichia coli transposon Tn10) and a cell membrane transducing peptide, is able to regulate transcription from a tetracycline responsive promoter (pCMV2xtetO2). When added directly to cultured cells, TetR fused to the full-length Antennapedia homeodomain (AntpHD) from Drosophila (TetRAntp), was able to selectively repress transcription in cells transiently transfected with a tetracycline-regulated reporter transcription unit. Moreover, TetRAntp could repress expression of a tetracycline responsive reporter transcription unit stably integrated into the genome of HeLa cells, demonstrating the possibility of manipulating endogenous gene expression by cell-permeable transcription factors.


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