Artificial Ribozyme Switches Containing Natural Riboswitch Aptamer Domains

Abstract

RNA Lego: The use of natural riboswitch aptamers in synthetic RNA switches (see picture) should broaden the scope of artificial RNA regulators dramatically. It is shown that thiamine pyrophosphate (TPP) aptamers can be used in engineered devices as very sensitive switches of gene expression in unmodified organisms. The approach demonstrates that intrinsic metabolites can be utilized as external effectors of cellular functions.

A Matter of Location: Influence of G-Quadruplexes on Escherichia coli Gene Expression

“…Different systems have been described using engineered devices as sensitive switches of gene expression in prokaryotic organisms (Klauser and Hartig, 2013;Wieland et al, 2009;Wieland and Hartig, 2007;Winkler et al, 2002). In this study, we successfully constructed a system where the SD region was masked via a hairpin structure.…”

RNA-mediated regulation in bacteria: from natural to artificial systems

“…In this case, the ribozyme cleaves the mRNA close to the SD, and in turn regulates translation [79]. Jin et al have recently constructed a theophyllin-responsive riboswitch upstream the ribosome binding site of E. coli csrA mRNA to express the essential protein from low to high levels.…”

Small‐Molecule‐Dependent Regulation of Transfer RNA in Bacteria

“…We have successfully used this "gain of function" principle of cleavage fragment dissociation before in order to liberate the ribosome binding site for switching on mRNA translation. [4][5][6] Indeed, when we connected an active HHR to the tRNA Ser CUA , strong eGFP expression occurred rivalling the level of wt-eGFP mRNA expression in combination with an unaltered tRNA (Figure 1 c, HHR-tRNA fusion). The fact that the combination of a suppressor tRNA with the corresponding amber mRNA results in expression levels of the reporter comparable to the natural system is intriguing since usually amber suppression is known to occur with maximum efficiencies of around 20-30 %.…”

“…[2,3] Recently, Ogawa and Maeda as well as our own group have introduced a different strategy by utilizing ligand-dependent ribozymes in order to switch on or off the translation of a given mRNA in bacteria. [4][5][6][7] Apart from these hammerhead-based systems for controlling mRNA translation in bacteria, ligand-dependent ribozymes have been utilized in yeast to program RNA-based Boolean logic gates. [8] In our opinion, the use of ligand-dependent, selfcleaving ribozymes is advantageous since it can be generalized for controlling RNA classes other than mRNAs.…”