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A novel positive tetracycline-dependent transactivator (rtTA) variant with reduced background activity and enhanced
Martin R Kämper1, Gabi Gohla, Gregor Schlüter
1Institute of Human Genetics, University of Göttingen, Göttingen, Germany.
Abstract:
The tetracycline-controlled transcription system has become one of the most potent systems for experimental manipulations of transcription levels in vivo. Here we report on rtTA variants, which were generated by combining the existing positively regulated Tet repressor domains of rtTA and rtTA-M2 with a modified and multimerized minimal transactivation domain from VP16 (L-domain). A transactivator with multimerized L-domains shows drastically reduced background activity and enhanced transcriptional activation on different tetracycline-responsive promoters. The new rtTA variants require higher doses of doxycycline and display a more linear dose-response curve than the original rtTA or rtTA-M2 proteins.