Monitoring the expression level of coding and non-coding RNAs using a TetR inducing aptamer tag.

RNA aptamers have been widely used as regulators for conditional gene expression. The TetR binding aptamer can activate tetracycline repressor TetR controlled gene expression with high efficiency. Here we demonstrate that the aptamer can also activate TetR controlled gene expression when expressed in the context of a natural transcripts. The aptamer was inserted into the untranslated regions of mRNAs as well as into small non-coding RNAs and was expressed both from a plasmid and from an endogenous locus. Our data suggest that the aptamer is a valuable tool to easily monitor the expression level of different RNAs, and it therefore represents a powerful tool for the construction of complex synthetic gene networks

An RNA aptamer that induces transcription.

We identified an RNA aptamer that induces TetR-controlled gene expression in Escherichia coli when expressed in the cell. The aptamer was found by a combined approach of in vitro selection for TetR binding and in vivo screening for TetR induction. The smallest active aptamer folds into a stem-loop with an internal loop interrupting the stem. Mutational analysis in vivo and in-line probing in vitro reveal this loop to be the protein binding site. The TetR-inducing activity of the aptamer directly correlates with its stability and the best construct is as efficient as the natural inducer tetracycline. Because of its small size, high induction efficiency, and the stability of the TetR aptamer under in vivo conditions, it is well suited to be an alternative RNA-based inducer of TetR-controlled gene expression