Designing Synthetic Flexible Gene Regulation Networks Using RNA Devices in Cyanobacteria

In recent years, studies on the development of gene regulation tools in cyanobacteria have been extensively conducted toward efficient production of valuable chemicals. However, there is considerable scope for improving the economic feasibility of production. To improve a recently reported gene induction system using anhydrotetracycline (aTc)–TetR and an endogenous gene repression system using small antisense RNA in the filamentous nitrogen-fixing cyanobacterium Anabaena sp. PCC 7120 (Anabaena), we constructed a positive feedback loop, in which gfp and a small antisense RNA for tetR are controlled by an aTc-inducible promoter. GFP expression in this improved system was higher and longer than the system lacking tetR repression. In addition, by using TetR aptamer and a riboswitch, we succeeded in achieving a superior and longer induction of GFP expression even under high-light conditions. Hence, efficient gene induction systems were established in Anabaena by designing a gene regulation network using RNA-based tools.