Cyanobacteria are ideal cellular factories for biochemical production because of their ability to fix CO2 by photosynthesis and convert this molecule into biochemicals. Previously, we engineered a riboregulator that enables post-transcriptional gene regulation in the cyanobacterium Synechocystis sp. PCC 6803. Here, we improved the riboregulator by designing two RNA species, taRNA and crRNA, to enhance its induction fold. We inserted nucleotides into the crRNA loop to enhance intermolecular hybridization and successfully improved its induction fold. The engineered riboregulator exhibited a higher induction fold than the previously engineered riboregulator in both Escherichia coli and Synechocystis sp. PCC 6803. This improved riboregulator can be used to control gene expression over a wide dynamic range in cyanobacteria.
Keywords: Cyanobacteria; RNA engineering; post-transcriptional gene regulation; riboregulator; synthetic biology.