Development of an RNA detection system using bioluminescence resonance energy transfer

Here we demonstrate a new RNA detection system using bioluminescence resonance energy transfer (BRET). Recombinant proteins were constructed consisting of enhanced yellow fluorescence protein (EYFP) and Renilla luciferase (RLuc) tethered by Arginine Rich Motif (ARM) peptides, which recognize specific RNA motifs and change their conformation upon binding to them. Conformational transitions of ARM peptides upon RNA binding then trigger a change in the BRET signal between RLuc and EYFP. Additionally, in order to detect arbitrarily selected RNAs, split-RNA probes were designed to reconstruct the ARM peptide binding motif by hybridizing with the target RNA. Combination of BRET probes and split-RNA probes enables to choose and detect target RNA sequences. (C) 2010 Elsevier B.V. All rights reserved.