title: A rapid, sensitive, and selective bioluminescence resonance energy transfer (BRET)-based nucleic acid sensing system.
abstract: Here we report the design of a bioluminescence resonance energy transfer (BRET)-based sensing system that could detect nucleic acid target in 5 min with high sensitivity and selectivity. The sensing system is based on adjacent binding of oligonucleotide probes labeled with Renilla luciferase (Rluc) and quantum dot (Qd) on the nucleic acid target. Here Rluc, a bioluminescent protein that generates light by a chemical reaction, is employed as an energy donor, and a quantum dot is used as an energy acceptor. Bioluminescence emission of Rluc overlaps with the Qd absorption whereas the emission of Qd is shifted from the emission of Rluc allowing for monitoring of BRET. In the presence of target, the labeled probes bind adjacently in a head-to-head fashion leading to BRET from Rluc to Qd upon addition of a substrate coelenterazine. The sensing system could detect target nucleic acid in buffer as well as in Escherichia coli cellular matrix in 5 min with a detection limit of 0.54 pmol. The ability to detect target nucleic acid rapidly in a cellular matrix with high sensitivity will prove highly beneficial in biomedical and environmental applications.
authors.affiliation: Department of Biochemistry and Molecular Biology, Miller School of Medicine, University of Miami, Miami, FL 33136, USA.
authors.forename: Manoj
authors.initials: M
authors.lastname: Kumar
authors.forename: Daohong
authors.initials: D
authors.lastname: Zhang
authors.forename: David
authors.initials: D
authors.lastname: Broyles
authors.forename: Sapna K
authors.initials: SK
authors.lastname: Deo
mesh_terms: D060807:Bioluminescence Resonance Energy Transfer Techniques / Q000295:instrumentation*; D015374:Biosensing Techniques / Q000295:instrumentation*; D004867:Equipment Design; D019544:Equipment Failure Analysis; D009696:Nucleic Acids / Q000032:analysis*; D012680:Sensitivity and Specificity
chemical_list: D009696:Nucleic Acids