High-Resolution Single-Molecule FRET via DNA eXchange (FRET X)
M. Filius (TU Delft - BN/Chirlmin Joo Lab, Kavli institute of nanoscience Delft)
Sung Hyun Kim (TU Delft - BN/Chirlmin Joo Lab, Kavli institute of nanoscience Delft)
Ivo Severins (Kavli institute of nanoscience Delft, TU Delft - BN/Chirlmin Joo Lab)
C. Joo (TU Delft - BN/Chirlmin Joo Lab, Kavli institute of nanoscience Delft)
More Info
expand_more
Other than for strictly personal use, it is not permitted to download, forward or distribute the text or part of it, without the consent of the author(s) and/or copyright holder(s), unless the work is under an open content license such as Creative Commons.
Abstract
Single-molecule FRET is a versatile tool to study nucleic acids and proteins at the nanometer scale. However, currently, only a couple of FRET pairs can be reliably measured on a single object, which makes it difficult to apply single-molecule FRET for structural analysis of biomolecules. Here, we present an approach that allows for the determination of multiple distances between FRET pairs in a single object. We use programmable, transient binding between short DNA strands to resolve the FRET efficiency of multiple fluorophore pairs. By allowing only a single FRET pair to be formed at a time, we can determine the pair distance with subnanometer precision. The distance between other pairs are determined by sequentially exchanging DNA strands. We name this multiplexing approach FRET X for FRET via DNA eXchange. Our FRET X technology will be a tool for the high-resolution analysis of biomolecules and nanostructures.
help_outline