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Cascade Amplification-Mediated In Situ Hot-Spot Assembly for MicroRNA Detection and Molecular Logic Gate Operations
journal contribution
posted on 2018-03-23, 19:33 authored by Sha Yu, Yingying Wang, Li-Ping Jiang, Sai Bi, Jun-Jie ZhuMicroRNAs (miRNAs) play important
roles in many biological processes
and are associated with various diseases, especially cancers. Combination
of technological developments such as nanomaterials, functional enzyme-mediated
reactions, and DNA nanotechnology holds great potential for high-performance
detection of miRNAs in molecular diagnostic systems. In this work,
we have fabricated a cascade signal amplification platform through
integrating duplex-specific nuclease (DSN)-assisted target recycling
with catalytic hairpin assembly (CHA) reaction for the detection of
microRNA-141 (miR-141). The target recycling process driven by DSN
results in highly amplified translation of target miRNA to single-stranded
connector DNA fragments. The CHA reaction is further initiated by
connector DNAs using hairpin-modified gold nanoparticles (HP-AuNPs)
as the sensing unit, leading to the formation of AuNP network architecture
on the electrode for electrochemical and photoelectrochemical detection
of miR-141 in signal-on and signal-off modes, respectively. The developed
electrochemical biosensor exhibits a detection limit down to 25.1
aM miR-141 (60 copies in 4 μL sample) and excellent selectivity
to discriminate a single base-mismatched sequence and other miRNAs.
This assay is also applied to the determination of miR-141 in total
RNAs extracted from human breast cancer cells (MDA-MB-231), confirming
the applicability of this method for absolute quantification of specific
miRNAs in real-world samples. Furthermore, two-input AND and INHIBIT
(INH) logic gates are constructed to detect miRNAs. In particular,
the AND gate achieves cell-specific gate activation based on expression
profiles of miR-141 and microRNA-21 (miR-21). Therefore, our proposed
cascade amplification platform has great potential applications in
miRNA-related clinical diagnostics and biochemical research.
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Keywords
AuNP network architectureCHAdetectionMDA-MBSitu Hot-Spot AssemblyDSNcell-specific gate activationhairpin-modified gold nanoparticlesINHbreast cancer cellsmiR -1414 μ L samplemiRNAMolecular Logic Gate Operations MicroRNAselectrochemical biosensor exhibitsINHIBITtarget recycling processcascade amplification platformsingle-stranded connector DNA fragmentscascade signal amplification platform
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