Engineering Naringenin-Specific Artificial Riboswitches through Evolutionary approach

Abstract

Various riboswitch-based genetic devices have been developed for diverse applications. While rational approaches have been used to redesign riboswitches to achieve distinct dose-response curves, this method is limited in its applicability. To address this, we developed an evolutionary approach for riboswitch engineering. Initially, we created a library of naringenin (NRN) aptamers through SELEX and constructed an NRN riboswitch library incorporating these aptamers. Enrichments were then performed through dual selection under medium and high NRN concentrations. Following selection, we characterized the dose-response curves of the enriched NRN riboswitches, classifying them based on their half maximal effective concentration relative to the NRN concentration during enrichment.