Rational development of FMN-based orthogonal riboswitch that functions in response to specific non-cognate ligand.

ABSTRACT: Re-engineering natural riboswitches into orthogonal RNA switches by making them functional in response to exogenous ligands but unresponsive to endogenous cognate ligands is a promising yet less explored strategy for developing gene regulatory tools. Herein, we rationally engineer the aptamer domain of one of the largest and biotechnologically relevant flavin mononucleotide (FMN) riboswitch class, which specifically binds to synthetic ligands with a high binding affinity (K_D = ~ 54-75 nM) and regulates gene expression in vitro, in prokaryotic, and eukaryotic system, while being unresponsive to FMN. To develop the orthogonal aptamers, we rationally alter key tertiary interactions, such as A/G minor motifs and base triples located in the periphery of the FMN binding pocket. The bi

SUBMITTER: Ojha D 

PROVIDER: S-EPMC12219259 | biostudies-literature | 2025 Jul

REPOSITORIES: biostudies-literature