Rational design of a synthetic mammalian riboswitch as a ligand-responsive -1 ribosomal frame-shifting stimulator

Lin, Ya-Hui; Chang, Kung-Yao

In this study, we take advantage of an extra stem-loop of SARS-CoV −1 PRF stimulation pseudoknot to show that this stem-loop can be replaced by an RNA aptamer to design a ligand-responsive −1 PRF stimulator with activity that rivals those of viral and metabolite-responsive stimulators. We further demonstrate the in vitro improvement of ligand responsiveness and function of the engineered riboswitch as a ligand-responsive −1 PRF stimulator in a stable human cell line. Thus, this scaffold should make a repertoire of RNA aptamers available for artificial riboswitch construction in ligand-dependent −1 PRF regulation of higher eukaryotes.