Tetracycline-controlled transcription in eukaryotes: novel transactivators with graded transactivation potential.
Several tetracycline-controlled transactivators (tTA) were generated which differ in their activation potential by >3 orders of magnitude. The transactivators are fusions between the Tet repressor and minimal transcriptional activation domains derived from Herpes simplex virus protein 16 (VP16). By reducing the VP16 moiety of the previously described tTA to 12 amino acids, potential targets for interactions with various cellular transcription factors were eliminated, as were potential epitopes which may elicit a cellular immune response. When compared with the originally described tTA, these new transactivators are tolerated at higher intracellular concentrations. This will facilitate establishment of tet regulatory systems under a variety of conditions, but particularly when cell type-restricted tetracycline-controlled gene expression is to be achieved in transgenic organisms via homologous recombination.
Baron
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U
ZMBH, Im Neuenheimer Feld 282, 69120 Heidelberg, Germany.
Gossen
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Bujard
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H
D001426:Bacterial Proteins / Q000235:genetics; D002352:Carrier Proteins; D005057:Eukaryotic Cells; D006367:HeLa Cells; D017405:Herpes Simplex Virus Protein Vmw65 / Q000235:genetics; D006801:Humans; D008156:Luciferases / Q000378:metabolism; D011993:Recombinant Fusion Proteins / Q000235:genetics; D012097:Repressor Proteins / Q000235:genetics; D013752:Tetracycline / Q000494:pharmacology*; D015534:Trans-Activators / Q000187:drug effects* / Q000378:metabolism; D014158:Transcription, Genetic / Q000187:drug effects*; D015533:Transcriptional Activation
D001426:Bacterial Proteins; D002352:Carrier Proteins; D017405:Herpes Simplex Virus Protein Vmw65; D011993:Recombinant Fusion Proteins; D012097:Repressor Proteins; C083139:Tet O resistance protein, Bacteria; D015534:Trans-Activators; C034811:tetracycline resistance-encoding transposon repressor protein; D008156:Luciferases; D013752:Tetracycline