Spectroscopic Features of Dual Fluorescence/Luminescence Resonance Energy-Transfer Molecular Beacons
Citations Over TimeTop 10% of 2003 papers
Abstract
Molecular beacons have the potential to become a powerful tool in gene detection and quantification in living cells. Here we report a novel dual molecular beacons approach to reduce false-positive signals in detecting target nucleic acids in homogeneous assays. A pair of molecular beacons, each containing a fluorescence quencher and a reporter fluorophore, one with a donor and a second with an acceptor fluorophore, hybridize to adjacent regions on the same target resulting in fluorescence resonance energy transfer (FRET). The detection of a FRET signal leads to a substantially increased signal-to-background ratio compared with that seen in single molecular beacon assays and enables discrimination between fluorescence due to specific probe/target hybridization and a variety of possible false-positive events. Further, when a lanthanide chelate is used as a donor in a dual-probe assay, extremely high signal-to-background ratios can be achieved owing to the long lifetime and sharp emission peaks of the donor and the time-gated detection of acceptor fluorescence emission. These new approaches allow for the ultrasensitive detection of target molecules in a way that could be readily applied to real-time imaging of gene expression in living cells.
Related Papers
- → Spectroscopic Features of Dual Fluorescence/Luminescence Resonance Energy-Transfer Molecular Beacons(2003)101 cited
- → Fluorescent proteins for single‐molecule fluorescence applications(2008)72 cited
- → A highly selective and sensitive ratiometric fluorescent probe for pH measurement based on fluorescence resonance energy transfer(2015)8 cited
- → Introduction of a Highly Photodurable and Common-laser Excitable Fluorescent Amino Acid into a Peptide as a FRET Acceptor for Protease Cleavage Detection(2010)9 cited
- → Genotyping by Guanosine-Dependent Quenching of Single-Labeled Fluorescein Probes(2002)