Key result
Adding 2,4-dinitrotoluene to riboswitch-engineered E. coli increases fluorescence ~10-fold vs. no target.
Why the study?
A novel riboswitch responsive to 2,4-dinitrotoluene (DNT) was engineered to regulate protein translation in E. coli cells.
Effect estimate: 10-fold increase
May enable bacterial biosensors for explosives; leaves open validation before any clinical or cardiovascular translation.
Riboswitches are RNA sequences that regulate expression of associated downstream genes in response to the presence or absence of specific small molecules. A novel riboswitch that activates protein translation in E. coli cells in response to 2,4-dinitrotoluene (DNT) has been engineered. A plasmid library was constructed by incorporation of 30 degenerate bases between a previously described trinitrotoluene aptamer and the ribosome binding site. Screening was performed by placing the riboswitch library upstream of the Tobacco Etch Virus (TEV) protease coding sequence in one plasmid; a second plasmid encoded a FRET-based construct linked with a peptide containing the TEV protease cleavage site. Addition of DNT to bacterial culture activated the riboswitch, initiating translation of TEV protease. In turn, the protease cleaved the linker in the FRET-based fusion protein, causing a change in fluorescence. This new riboswitch exhibited a 10-fold increase in fluorescence in the presence of 0.5 mM DNT compared to the system without target.
No takes yet. Share an insight, caveat, or question.
Davidson et al. (2012) studied this question. 2,4-dinitrotoluene (DNT) vs. system without target was evaluated on Fluorescence change (10-fold increase). Addition of 0.5 mM 2,4-dinitrotoluene to E. coli cells engineered with a novel synthetic riboswitch resulted in a 10-fold increase in fluorescence compared to the system without target.
Synapse has enriched 5 closely related papers on similar clinical questions. Consider them for comparative context: