Improvement of the reverse tetracycline transactivator by single amino acid substitutions that reduce leaky target gene expression to undetectable levels

• DOI: 10.1038/srep27697
• Publication Date: 2016-06-21T09:57:34Z
• Authors: Ian J. Roney; Adam D. Rudner; Jean-François Cou...; Mads Kærn
• Keywords: Inducer; Transcription

Conditional gene expression systems that enable inducible and reversible transcriptional control are essential research tools have broad applications in biomedicine biotechnology. The reverse tetracycline activator is a canonical system for engineered enables graded gratuitous modulation of target transcription eukaryotes from yeast to human cell lines transgenic animals. However, the has tendency activate even absence this leaky impedes its use. Here, we identify single amino-acid substitutions greatly enhance dynamic range by reducing undetectable levels while retaining high capacity presence inducer. While mutations increase inducer concentration required full induction, additional sensitivity-enhancing can compensate effect confer degree robustness system. novel transactivator variants will be useful where tight tunable regulation paramount.