Light-controlled regulation of eukaryotic gene expression using a diffusible effector

This project focuses on the design of a small synthetic, trans-acting noncoding RNA to directly regulate gene expression by light irradiation in a eukaryotic system at the mRNA level. This trans-acting noncoding RNA is designed to incorporate an in vitro-selected small ligand-binding domain, called an aptamer domain. This aptamer domain selectively binds to one but not the other of the two isomers of a dihydropyrene photo-switch compound. Such an aptamer sequence allowed us to design an allosteric riboregulator, a RNA molecule with a regulatory function whose mode of activity depends on its photo-switch ligand binding to the aptamer domain, which in turn is influenced by light irradiation at an appropriate wavelength. This light-controlled riboregulator is referred to as an antiswitch. This antiswitch is specifically designed to control the expression of Green Fluorescent protein (GFP) in vivo in Saccharomyces cerevisiae as a model organism.