西亚试剂：Versatile RNA-sensing transcriptional regulators for engine

Versatile RNA-sensing transcriptional regulators for engineering genetic networks

Lucks, Julius B.; Qi, Lei; Mutalik, Vivek K.; Wang, Denise; Arkin, Adam P.

The widespread natural ability of RNA to sense small molecules and regulate genes has become an important tool for syntheticbiology in applications as diverse as environmental sensing and metabolic engineering. Previous work in RNA synthetic biologyhas engineered RNA mechanisms that independently regulate multiple targets and integrate regulatory signals. However, intracellularregulatory networks built with these systems have required proteins to propagate regulatory signals. In this work, we removethis requirement and expand the RNA synthetic biology toolkit by engineering three unique features of the plasmid pT181 antisense-RNA-mediatedtranscription attenuation mechanism. First, because the antisense RNA mechanism relies on RNA-RNA interactions, we show howthe specificity of the natural system can be engineered to create variants that independently regulate multiple targets inthe same cell. Second, because the pT181 mechanism controls transcription, we show how independently acting variants can beconfigured in tandem to integrate regulatory signals and perform genetic logic. Finally, because both the input and outputof the attenuator is RNA, we show how these variants can be configured to directly propagate RNA regulatory signals by constructingan RNA-meditated transcriptional cascade. The combination of these three features within a single RNA-based regulatory mechanismhas the potential to simplify the design and construction of genetic networks by directly propagating signals as RNA molecules.