| DOI | Trouver le DOI : https://doi.org/10.1016/j.ab.2011.01.030 |
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| Auteur | Rechercher : Nasheri, N.1; Rechercher : Cheng, J.1; Rechercher : Singaravelu, R.1; Rechercher : Wu, P.; Rechercher : McDermott, M.T.; Rechercher : Pezacki, J.P.1 |
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| Affiliation | - Conseil national de recherches du Canada. Institut Steacie des sciences moléculaires du CNRC
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| Format | Texte, Article |
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| Sujet | 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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| Résumé | 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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| Date de publication | 2011 |
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| Dans | |
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| Langue | anglais |
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| Publications évaluées par des pairs | Oui |
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| Numéro NPARC | 21271288 |
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| Exporter la notice | Exporter en format RIS |
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| Signaler une correction | Signaler une correction (s'ouvre dans un nouvel onglet) |
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| Identificateur de l’enregistrement | e53bbd7a-52b5-4d50-b6a3-9b3a6742263d |
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| Enregistrement créé | 2014-03-24 |
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| Enregistrement modifié | 2020-04-21 |
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