DOI | Resolve DOI: https://doi.org/10.1016/j.ab.2011.01.030 |
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Author | Search for: Nasheri, N.1; Search for: Cheng, J.1; Search for: Singaravelu, R.1; Search for: Wu, P.; Search for: McDermott, M.T.; Search for: Pezacki, J.P.1 |
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Affiliation | - National Research Council of Canada. NRC Steacie Institute for Molecular Sciences
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Format | Text, Article |
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Subject | gold; microRNA; oligonucleotide; protein p19; RNA; enzyme assay; enzyme immunoassay; enzyme linked immunosorbent assay; genetic procedures; hepatoma cell; human cell; molecular size; nucleic acid immobilization; protein binding; protein expression; protein purification; RNA analysis; RNA extraction; RNA interference; RNA isolation; RNA probe; sensitivity analysis; surface plasmon resonance; Tombusvirus; Adsorption; Base Pair Mismatch; Cell Line, Tumor; Electrophoretic Mobility Shift Assay; Enzyme Assays; Genes, Suppressor; Immunoassay; Limit of Detection; Magnetics; MicroRNAs; RNA Interference; RNA Probes; Surface Plasmon Resonance; Tombusvirus; Viral Proteins; Metazoa; Tombusvirus |
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Abstract | MicroRNAs (miRNAs) are endogenous posttranscriptional regulators found in all metazoa and play crucial roles in virtually all cellular processes. Their aberrant expression has been linked to several diseased states; therefore, techniques capable of sensitive and specific profiling of the miRNA milieu will have significant application in prognostics, diagnostics, and therapeutics. Here we present a method for rapid quantification of miRNA levels using p19, a tombusvirus-encoded suppressor of RNA interference with sequence-independent and size-selective affinity toward 19-bp RNA duplexes. We present a surface plasmon resonance (SPR)-based miRNA sensing method where RNA probes are immobilized on gold surfaces demonstrating p19's utility in recognition of miRNA-bound probes. This allows detection of miRNAs in the low nanomolar range. To increase the sensitivity, a bead-based enzyme immunoassay was performed, and this technique displays a lower detection limit of 1 fmol and a linear dynamic range from 1 pmol to 1 fmol. © 2011 Published by Elsevier Inc. All rights reserved. |
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Publication date | 2011 |
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In | |
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Language | English |
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Peer reviewed | Yes |
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NPARC number | 21271288 |
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Export citation | Export as RIS |
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Report a correction | Report a correction (opens in a new tab) |
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Record identifier | e53bbd7a-52b5-4d50-b6a3-9b3a6742263d |
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Record created | 2014-03-24 |
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Record modified | 2020-04-21 |
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