Intermolecular dark resonance energy transfer (DRET): Upgrading fluorogenic DNA sensing
Artikel i vetenskaplig tidskrift, 2021

The sensitivity of FRET-based sensing is usually limited by the spectral overlaps of the FRET donor and acceptor, which generate a poor signal-to-noise ratio. To overcome this limitation, a quenched donor presenting a large Stokes shift can be combined with a bright acceptor to perform Dark Resonance Energy Transfer (DRET). The consequent fluorogenic response from the acceptor considerably improves the signal-to-noise ratio. To date, DRET has mainly relied on a donor that is covalently bound to the acceptor. In this context, our aim was to develop the first intermolecular DRET pair for specific sensing of nucleic acid sequences. To this end, we designed DFK, a push-pull probe based on a fluorenyl π-platform that is strongly quenched in water. DFK was incorporated into a series of oligonucleotides and used as a DRET donor with Cy5-labeled complementary sequences. In line with our expectations, excitation of the dark donor in the double-labeled duplex switched on the far-red Cy5 emission and remained free of cross-excitation. The DRET mechanism was supported by time-resolved fluorescence measurements. This concept was then applied with binary probes, which confirmed the distance dependence of DRET as well as its potency in detecting sequences of interest with low background noise.


Guillaume Barnoin


Janah Shaya


Khalifa University

Ludovic Richert

Université de Strasbourg

Hoang Ngoan Le


Chalmers, Kemi och kemiteknik, Kemi och biokemi

Steve Vincent


Vincent Guérineau

Université Paris-Saclay

Yves Mely

Université de Strasbourg

Benoît Y. Michel


Alain Burger


Nucleic Acids Research

0305-1048 (ISSN) 1362-4962 (eISSN)

Vol. 49 12 e72


Atom- och molekylfysik och optik


Annan elektroteknik och elektronik





Mer information

Senast uppdaterat