Suppression of gene expression by a cell-permeable Tet repressor.
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.
Mortlock
Alison
A
Biogeny PLC and Department of Biology and Biochemistry, SAF Building, Imperial College, London SW7 2AZ, UK.
Low
Walter
W
Crisanti
Andrea
A
D000818:Animals; D051943:Antennapedia Homeodomain Protein; D002463:Cell Membrane Permeability; D029721:Drosophila Proteins; D004331:Drosophila melanogaster; D004926:Escherichia coli / Q000235:genetics; D005786:Gene Expression Regulation / Q000187:drug effects*; D017930:Genes, Reporter / Q000235:genetics; D006367:HeLa Cells; D018398:Homeodomain Proteins / Q000235:genetics / Q000378:metabolism; D006801:Humans; D009687:Nuclear Proteins; D009876:Operon / Q000235:genetics; D011401:Promoter Regions, Genetic / Q000235:genetics; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism; D012097:Repressor Proteins / Q000235:genetics / Q000378:metabolism*; D013752:Tetracycline / Q000494:pharmacology*; D013753:Tetracycline Resistance / Q000235:genetics; D014157:Transcription Factors; D014162:Transfection
D051943:Antennapedia Homeodomain Protein; C497974:Antp protein, Drosophila; D029721:Drosophila Proteins; D018398:Homeodomain Proteins; D009687:Nuclear Proteins; D011993:Recombinant Fusion Proteins; D012097:Repressor Proteins; D014157:Transcription Factors; C034811:tetracycline resistance-encoding transposon repressor protein; D013752:Tetracycline