Single-Molecule FRET Detection of Sub-Nanometer Distance Changes in the Range below a 3-Nanometer Scale

Single-molecule fluorescence energy transfer (FRET) detection has become a key technique to monitor intra- and intermolecular distance changes in biological processes. As the sensitive detection range of conventional FRET pairs is limited to 3-8 nm, complement probes are necessary for extending this...

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Bibliographic Details
Published in:Biosensors (Basel) 2020-11, Vol.10 (11), p.168
Main Authors: Son, Heyjin, Mo, Woori, Park, Jaeil, Lee, Joong-Wook, Lee, Sanghwa
Format: Article
Language:English
Subjects:
Communication
conformational dynamics
DNA
Fluorescence
Fluorescence Resonance Energy Transfer
FRET probe
Nanotechnology
single-molecule FRET
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Summary:Single-molecule fluorescence energy transfer (FRET) detection has become a key technique to monitor intra- and intermolecular distance changes in biological processes. As the sensitive detection range of conventional FRET pairs is limited to 3-8 nm, complement probes are necessary for extending this typical working range. Here, we realized a single-molecule FRET assay for a short distance range of below 3 nm by using a Cy2-Cy7 pair having extremely small spectral overlap. Using two DNA duplexes with a small difference in the labeling position, we demonstrated that our assay can observe subtle changes at a short distance range. High sensitivity in the range of 1-3 nm and compatibility with the conventional FRET assay make this approach useful for understanding dynamics at a short distance.
ISSN:2079-6374
2079-6374
DOI:10.3390/bios10110168