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2. Development of an RNA detection system using bioluminescence resonance energy transfer

Author

Andou, T., Endoh, T., mie, masayasu, and KOBATAKE, EIRY

Subjects
Chemistry, Energy transfer, Metals and Alloys, Resonance, RNA, Condensed Matter Physics, Fluorescence, Surfaces, Coatings and Films, Electronic, Optical and Magnetic Materials, law.invention, RNA Motifs, Biochemistry, Peptide binding motif, law, Materials Chemistry, Biophysics, Recombinant DNA, Bioluminescence, Electrical and Electronic Engineering, Instrumentation
Abstract

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 A rginine R ich M otif (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.

Published
2011

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