Kinetic FRET Assay to Measure Binding-Induced Conformational Changes of Nucleic Acids.

ABSTRACT: The interaction of small molecules or proteins with RNA or DNA often involves changes in the nucleic acid (NA) folding and structure. A biophysical characterization of these processes helps us to understand the underlying molecular mechanisms. Here, we propose kinFRET (kinetics Förster resonance energy transfer), a real-time ensemble FRET methodology to measure binding and folding kinetics. With kinFRET, the kinetics of conformational changes of NAs (DNA or RNA) upon analyte binding can be directly followed via a FRET signal using a chip-based biosensor. We demonstrate the utility of this approach with two representative examples. First, we monitored the conformational changes of different formats of an aptamer (MN19) upon interaction with small-molecule analytes. Second, we characterized

SUBMITTER: Higuera-Rodriguez RA 

PROVIDER: S-EPMC10749467 | biostudies-literature | 2023 Dec

REPOSITORIES: biostudies-literature