title: Detection of miRNA using a double-strand displacement biosensor with a self-complementary fluorescent reporter.
abstract: Design of rapid, selective, and sensitive DNA and ribonucleic acid (RNA) biosensors capable of minimizing false positives from nuclease degradation is crucial for translational research and clinical diagnostics. We present proof-of-principle studies of an innovative micro-ribonucleic acid (miRNA) reporter-probe biosensor that displaces a self-complementary reporter, while target miRNA binds to the probe. The freed reporter folds into a hairpin structure to induce a decrease in the fluorescent signal. The self-complementarity of the reporter facilitates the reduction of false positives from nuclease degradation. Nanomolar limits of detection and quantitation were capable with this proof-of-principle design. Detection of miRNA occurs within 10 min and does not require any additional hybridization, labeling, or rinsing steps. The potential for medical applications of the reporter-probe biosensor is demonstrated by selective detection of a cancer regulating microRNA, Lethal-7 (Let-7a). Mechanisms for transporting the biosensor across the cell membrane will be the focus of future work.
authors.affiliation: Department of Chemistry, Oregon State University , 153 Gilbert Hall, Corvallis, Oregon 97331, United States.
authors.forename: Nicholas E
authors.initials: NE
authors.lastname: Larkey
authors.forename: C Kyle
authors.initials: CK
authors.lastname: Almlie
authors.forename: Victoria
authors.initials: V
authors.lastname: Tran
authors.forename: Marianne
authors.initials: M
authors.lastname: Egan
authors.forename: Sean M
authors.initials: SM
authors.lastname: Burrows
mesh_terms: D001483:Base Sequence; D015374:Biosensing Techniques / Q000379:methods*; D004720:Endonucleases / Q000378:metabolism; D005456:Fluorescent Dyes / Q000032:analysis / Q000737:chemistry* / Q000378:metabolism; D007700:Kinetics; D035683:MicroRNAs / Q000032:analysis* / Q000737:chemistry / Q000235:genetics; D008958:Models, Molecular; D009690:Nucleic Acid Conformation; D009693:Nucleic Acid Hybridization; D015345:Oligonucleotide Probes / Q000032:analysis / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D013816:Thermodynamics
chemical_list: D005456:Fluorescent Dyes; D035683:MicroRNAs; D015345:Oligonucleotide Probes; D004720:Endonucleases