Fig. 5.

Trans-activating RNA library. (A) Cis-repressing RNA is designed to partially complement itself and sequestrate the ribosome binding site. (B) Structure of trans-activating RNA. (C) Interaction of trans-activating RNA with cis-repressing RNA relieves the sequestration and allows the recruitment of ribosome and translation initiation. (D) Synthetic overlapping oligos was annealed together and assembled into the XbaI and KpnI digested pCDM4 vector backbone. Box region shows the overlapping region and sequence flanked by XbaI and KpnI is the core of the degenerate trans-activating RNA. (E) Translational strength of trans-activating RNA library screened by green fluorescence protein. (F) Statistical distribution of gene expression activation across the engineered trans-activating RNA library.