Transcription Driven by Reversible Photocontrol of Hyperstable G-Quadruplexes

Abstract

G-quadruplexes occur in promoter regions, 5'-untranslated regions of mRNA and telomeric regions, and they function as regulatory elements for various key biological events, such as transcription, translation, and telomere elongation. As the stability of G-quadruplexes dramatically impacts these biological processes, controlling G-quadruplex stability via external stimuli such as light enables regulation of important biological phenomena with high spatial and temporal resolution. Here, we report a method for reversible photoregulation of transcription by controlling the stability of G-quadruplexes via cis- trans photoisomerization of photochromic nucleobase (PCN). Transcription was effectively inhibited when the PCN-modified G-quadruplex was in a hyperstable state, whereas transcription activity recovered markedly when the G-quadruplex changed to an unstable state induced by trans to cis PCN photoisomerization. Moreover, a reversibly photoactivatable plasmid was constructed by introducing PCN-modified G-quadruplexes downstream of the cytomegalovirus promoter of the pCS2 plasmid, which was used to demonstrate photoregulation of gene expression in zebrafish embryos.