Abstract
Quantification of circulating microRNAs (miRNAs) is of great interest because of their potentials as disease biomarkers. Currently, quantitative reverse transcription polymerase chain reaction (qRT-PCR) and microarray are considered mainstream techniques for miRNA identification and quantitation. However, these techniques are challenged by the low levels and wide dynamic range (from aM to nM) of miRNAs in a physiological sample, as well as the difficulty in the implementation in point-of-care settings. Here, we describe a one-step label-free electrochemical sensing technique by assembling a triple-stem DNA-redox probe structure on a gold microelectrode and introducing a reductant, tris(2-carboxyethyl) phosphine hydrochloride (TCEP) in the detection buffer solution to achieve ultrasensitive miRNAs detection with a detection limit of 0.1 fM.
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References
Dong H, Lei J, Ding L, Wen Y, Ju H, Zhang X (2013) MicroRNA: function, detection, and bioanalysis. Chem Rev 113:6207–6233
Lewis BP, Burge CB, Bartel DP (2005) Conserved seed pairing, often flanked by adenosines, indicates that thousands of human genes are microRNA targets. Cell 120:15–20
Cullen BR (2009) Viral and cellular messenger RNA targets of viral microRNAs. Nature 457:421–425
Laterza OF, Lim L, Garrett-Engele PW, Vlasakova K, Muniappa N, Tanaka WK, Johnson JM, Sina JF, Fare TL, Sistare FD, Glaab WE (2009) Plasma MicroRNAs as sensitive and specific biomarkers of tissue injury. Clin Chem 55:1977–1983
Hayes J, Peruzzi PP, Lawler S (2014) MicroRNAs in cancer: biomarkers, functions and therapy. Trends Mol Med 20:460–469
Wang K, Zhang S, Marzolf B, Troisch P, Brightman A, Hu Z (2009) Circulating microRNAs, potential biomarkers for drug-induced liver injury. Proc Natl Acad Sci U S A 106:4402–4407
Michel V, Yuan Z, Ramsubir S, Bakovic M (2006) Choline transport for phospholipid synthesis. Exp Biol Med 231:490–504
Klein D (2002) Quantification using real-time PCR technology: applications and limitations. Trends Mol Med 8:257–260
Lim LP (2003) The microRNAs of Caenorhabditis elegans. Genes Dev 17:991–1008
Liang Y, Ridzon D, Wong L, Chen C (2007) Characterization of microRNA expression profiles in normal human tissues. BMC Genomics 8:166
Liu CG, Calin GA, Meloon B, Gamllel N, Sevignani C, Ferracin M, Dumitru CD, Shimizu M, Zupo S, Dono M, Alder H, Bullrich F, Negrini M, Croce CM (2004) An oligonucleotide microchip for genome-wide microRNA profiling in human and mouse tissues. Proc Natl Acad Sci U S A 101:9740–9744
Lodes MJ, Caraballo M, Suciu D, Munro S, Kumar A, Anderson B (2009) Detection of cancer with serum miRNAs on an oligonucleotide microarray. PLoS One 4:e6229
Asaga S, Kuo C, Nguyen T, Terpenning M, Giuliano AE, Hoon DSB (2011) Direct serum assay for MicroRNA-21 concentrations in early and advanced breast cancer. Clin Chem 57:84–91
Zhi F, Cao X, Xie X, Wang B, Dong W, Gu W, Ling Y, Wang R, Yang Y, Liu Y (2013) Identification of circulating MicroRNAs as potential biomarkers for detecting acute myeloid leukemia. PLoS One 8:e56718
Gaur A, Jewell DA, Liang Y, Ridzon D, Moore JH, Chen C, Ambros VR, Israel MA (2007) Characterization of microRNA expression levels and their biological correlates in human cancer cell lines. Cancer Res 67:2456–2468
Zhang J, Li Z, Wang H, Wang Y, Jia H, Yan J (2011) Ultrasensitive quantification of mature microRNAs by real-time PCR based on ligation of a ribonucleotide-modified DNA probe. Chem Commun 47:9465–9467
Gao ZQ, Yang ZC (2006) Detection of microRNAs using electrocatalytic nanoparticle tags. Anal Chem 78:1470–1477
Labib M, Khan N, Ghobadloo SM, Cheng J, Pezacki JP, Berezovski MV (2013) Three-mode electrochemical sensing of ultralow MicroRNA levels. J Am Chem Soc 135:3027–3038
Labib M, Ghobadloo SM, Khan N, Kolpashchikov DM, Berezovski MV (2013) Four-way junction formation promoting ultrasensitive electrochemical detection of MicroRNA. Anal Chem 85:9422–9427
Labib M, Khan N, Berezovski MV (2015) Protein electrocatalysis for direct sensing of circulating MicroRNAs. Anal Chem 87:1395–1403
Wang T, Viennois E, Merlin D, Wang G (2015) Microelectrode miRNA sensors enabled by enzyme less electrochemical signal amplification. Anal Chem 87:8173–8180
White RJ, Plaxco KW (2010) Exploiting binding-induced changes in probe flexibility for the optimization of electrochemical biosensors. Anal Chem 82:73–76
Acknowledgment
This work was supported by grants from the Department of Veterans Affairs (Merit Award: BX002526 to D.M.) and the National Institute of Health of Diabetes and Digestive and Kidney by the Grants RO1-DK-071594 and RO1-DK-064711 (to D.M.).
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Wang, T., Wang, G., Merlin, D., Viennois, E. (2017). MiRNA Quantitation with Microelectrode Sensors Enabled by Enzymeless Electrochemical Signal Amplification. In: Dalmay, T. (eds) MicroRNA Detection and Target Identification. Methods in Molecular Biology, vol 1580. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-6866-4_17
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DOI: https://doi.org/10.1007/978-1-4939-6866-4_17
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