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Publication: farFRET - Extending the Range in Single-Molecule FRET Experiments beyond 10 nm

Single-molecule Förster resonance energy transfer (smFRET) has become a powerful nanoscopic tool in studies of biomolecular structures and nanoscale objects; however, conventional smFRET measurements are generally blind to distances above 10 nm thus impeding the study of long-distance phenomena. In the publication, a research team of the Schlierf lab reports the development of farFRET, a technique that extends the range in single-molecule FRET (smFRET) measurements beyond the 10 nm line by enhanced energy transfer using multiple acceptors. The team demonstrates that farFRET can be readily employed to quantify FRET efficiencies and conformational dynamics using double-stranded DNA molecules, RecA-filament formation on single-stranded DNA and Holliday junction dynamics. farFRET allows quantitative measurements of large biomolecular complexes and nanostructures thus bridging the remaining gap to superresolution microscopy.

 

This figure illustrates the principle of farFRET, a single-molecule (sm) technique that utilizes the enhancement in Förster resonance energy transfer (FRET) from multiple acceptors to break the 10 nm limit in smFRET experiments. Donor excitation energy is transferred to a bundle of four acceptors on double-stranded DNA. The presence of multiple acceptors increases the probability of energy transfer and thereby amplifies the observed FRET efficiency, enabling the observation of long-distance phenomena on individual molecules. The artistic representation of farFRET was created by Mario Avellaneda.

Original publication:

Georg Krainer, Andreas Hartmann, and Michael Schlierf: farFRET: Extending the Range in Single-Molecule FRET Experiments beyond 10 nm, DOI: 10.1021/acs.nanolett.5b01878             

To discover more, please follow the link to the journal article or contact the authors.

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