title: A novel TetR-regulating peptide turns off rtTA-mediated activation of gene expression.
abstract: Conditional regulation of gene expression is a powerful and indispensable method for analyzing gene function. The "Tet-On" system is a tool widely used for that purpose. Here, the transregulator rtTA mediates expression of a gene of interest after addition of the small molecule effector doxycycline. Although very effective in rapidly turning on gene expression, the system is hampered by the long half-life of doxycycline which makes shutting down gene expression rapidly very difficult to achieve. We isolated an rtTA-binding peptide by in vivo selection that acts as a doxycycline antagonist and leads to rapid and efficient shut down of rtTA-mediated reporter gene expression in a human cell line. This peptide represents the basis for novel effector molecules which complement the "Tet-system" by enabling the investigator to rapidly turn gene expression not just on at will, but now also off.
authors.affiliation: Lehrstuhl für Mikrobiologie, Department Biologie, Friedrich-Alexander-Universität Erlangen-Nürnberg, Erlangen, Germany.
authors.forename: Sebastian
authors.initials: S
authors.lastname: Schmidt
authors.affiliation: Lehrstuhl für Mikrobiologie, Department Biologie, Friedrich-Alexander-Universität Erlangen-Nürnberg, Erlangen, Germany.
authors.forename: Christian
authors.initials: C
authors.lastname: Berens
authors.affiliation: Lehrstuhl für Mikrobiologie, Department Biologie, Friedrich-Alexander-Universität Erlangen-Nürnberg, Erlangen, Germany.
authors.forename: Marcus
authors.initials: M
authors.lastname: Klotzsche
mesh_terms: D000595:Amino Acid Sequence; D002460:Cell Line; D004318:Doxycycline / Q000037:antagonists & inhibitors / Q000494:pharmacology; D017930:Genes, Reporter / Q000235:genetics; D006801:Humans; D008969:Molecular Sequence Data; D010455:Peptides / Q000737:chemistry / Q000494:pharmacology*; D012097:Repressor Proteins / Q000378:metabolism*; D015533:Transcriptional Activation / Q000187:drug effects*
chemical_list: D010455:Peptides; D012097:Repressor Proteins; C034811:tetracycline resistance-encoding transposon repressor protein; D004318:Doxycycline