Recent progress in synthetic biology and metabolic engineering approaches has enabled the engineering of microbes for production of high-value products with important medical and biotechnological applications. Synthetic riboregulators that utilize highly specific RNA-RNA interactions have been engineered to control posttranscriptional gene expression. Riboregulators consist of a cis-repressor RNA (crRNA) element designed upstream of the 5’-untransalated region (5’-UTR) of the messenger RNA (mRNA) of the desired gene and a trans-activating riboregulator element (taRNA) that can bind to the 5’-UTR of the target mRNA and regulate gene expression. The specificity and stability of RNA-RNA interactions, tunability and fast response of riboregulators can be exploited for providing a wide dynamic range of protein expression and to control multiple genes in an engineered pathway. In this study, we describe a synthetic biology approach using riboregulators to fine tune gene expression using reporter genes. Utilizing <i>E. coli </i>as a model organism, we have developed crRNA and taRNA elements to control the translation of mRNA of interest. Importantly, unlike previously reported riboregulators, we have shown that we can achieve upto ~200 fold difference in activation of genes even with much lower concentrations of riboregulators in cells. Our overall goal is to engineer a carbon and energy efficient pathway, which has multiple genes under the control of riboregulators and is fine-tuned for optimal biosynthesis of acetyl-CoA.

Riboregulators as tunable gene switches for post-transcriptional control of gene expression

249th National Meeting (2015)

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