Development of Ligand-Controlled Riboregulators in E. coli

After seeing the capabilities of RNA regulators of protein expression in nature, synthetic biologists adapted the technology. The difference in the transition from transcription to translation between eukaryotes and bacteria resulted in the necessity of different technologies for eukaryotes and bacteria. Currently eukaryotes have the ligand-controlled antiswitches, but bacteria have transcription factor-controlled riboregulators. By combining antiswitch and riboregulator technology, a ligand-controlled riboregulator could be developed; thus, allowing riboregulators to be capable of responding to a much greater variety of molecules. Using a technique adapted from PCR mutagenesis, I have been able to generate full length self-complementary DNA parts from shorter segments. Using BioBricks to clone the parts into E. coli, I am replicating the original riboregulator device. Before attempting to make the device ligand controlled, I had to make sure a functional original riboregulator could be generated by our methods. Flow cytometry will be used in the future to determine functionality and efficiency. If the methods work, a ligand-controlled riboregulator will be built by substituting the taRNA complement to the crRNA into the antisense stem. If the ligand-controlled riboregulator can be developed, it will have applications for generating bacteria biological sensors