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PLoS One. 2014 May 8;9(5):e96546. doi: 10.1371/journal.pone.0096546. eCollection 2014.

A novel TetR-regulating peptide turns off rtTA-mediated activation of gene expression.

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Lehrstuhl für Mikrobiologie, Department Biologie, Friedrich-Alexander-Universität Erlangen-Nürnberg, Erlangen, Germany.


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.

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