Dynamic Monitoring of MicroRNA–DNA Hybridization Using DNAase-Triggered Signal Amplification

Dynamically monitoring microRNA (miRNA)–DNA reactions is critical for elucidating various biological processes. However, traditional strategies fail to capture this dynamic event because the original targets are preamplified. In the present study, we developed an amplification-free strategy for real...

Full description

Saved in:
Bibliographic Details
Published in:Analytical chemistry (Washington) 2015-06, Vol.87 (12), p.6303-6310
Main Authors: Qiu, Xiaopei, Liu, Xing, Zhang, Wei, Zhang, Hong, Jiang, Tianlun, Fan, Dongli, Luo, Yang
Format: Article
Language:English
Subjects:
Amplification
Biosensors
Coating
Deoxyribonucleases - metabolism
Deoxyribonucleic acid
DNA
DNA - blood
DNA - genetics
DNA - metabolism
Dynamics
Graphene
Humans
Hybridization
MicroRNAs - blood
MicroRNAs - genetics
MicroRNAs - metabolism
Monitoring
Nucleic Acid Amplification Techniques
Nucleic Acid Hybridization
Ribonucleic acid
Ribonucleic acids
RNA
Strategy
Citations: Items that this one cites
Items that cite this one
Online Access:Get full text
Tags: Add Tag
No Tags, Be the first to tag this record!
Description
Summary:Dynamically monitoring microRNA (miRNA)–DNA reactions is critical for elucidating various biological processes. However, traditional strategies fail to capture this dynamic event because the original targets are preamplified. In the present study, we developed an amplification-free strategy for real-time monitoring of miRNA–DNA hybridization that integrates the advantages of both duplex-specific nuclease (DSN)-triggered signal amplification and single-stranded DNA probe coating facilitated by reduced graphene oxide. DSN-mediated miRNA recognition was found to consist of two phases: hybridization and hybridization cleavage. In the presence of miRNA and DSN, hybridization of a 22-mer miRNA–DNA could be completed within 7 min by observing the angle increase in a surface plasmon resonance (SPR) biosensor. The subsequent hybridization-cleavage process could be visualized as a gradual SPR angle decrease that occurred until all coated probes were hydrolyzed. In addition, for miRNA-21 detection, the proposed linear signal amplification assay demonstrated a sensitivity of 3 fM over a dynamic range of 5 orders of magnitude.
ISSN:0003-2700
1520-6882
DOI:10.1021/acs.analchem.5b01159