Electrophoretic mobility shift as a molecular beacon-based readout for miRNA detection.
Electrophoretic mobility
Gel electrophoresis
Molecular beacons
microRNA (miRNA)
Journal
Biosensors & bioelectronics
ISSN: 1873-4235
Titre abrégé: Biosens Bioelectron
Pays: England
ID NLM: 9001289
Informations de publication
Date de publication:
01 Oct 2021
01 Oct 2021
Historique:
received:
18
02
2021
revised:
27
04
2021
accepted:
03
05
2021
pubmed:
2
6
2021
medline:
22
6
2021
entrez:
1
6
2021
Statut:
ppublish
Résumé
MicroRNAs are short, non-coding RNA sequences involved in gene expression regulation. Quantification of miRNAs in biological fluids involves time consuming and laborious methods such as Northern blotting or PCR-based techniques. Molecular beacons (MB) are an attractive means for rapid detection of miRNAs, although the need for sophisticated readout methods limits their use in research and clinical settings. Here, we introduce a novel method based on delayed electrophoretic mobility, as a quantitative means for detection of miRNAs-MB hybridization. Upon hybridization with the target miRNAs, MB form a fluorescent duplex with reduced electrophoretic mobility, thus bypassing the need for additional staining. In addition to emission of light, the location of the fluorescent band on the gel acts as an orthogonal validation of the target identity, further confirming the specificity of binding. The limit of detection of this approach is approximately 100 pM, depending on the MB sequence. The method is sensitive enough to detect specific red blood cell miRNAs molecules in total RNA, with single nucleotide specificity. Altogether, we describe a rapid and affordable method that offers sensitive detection of single-stranded small DNA and RNA sequences.
Identifiants
pubmed: 34062334
pii: S0956-5663(21)00344-4
doi: 10.1016/j.bios.2021.113307
pmc: PMC8461749
mid: NIHMS1705286
pii:
doi:
Substances chimiques
MicroRNAs
0
Types de publication
Journal Article
Langues
eng
Sous-ensembles de citation
IM
Pagination
113307Subventions
Organisme : NCI NIH HHS
ID : R01 CA218500
Pays : United States
Organisme : NCATS NIH HHS
ID : UH3 TR002881
Pays : United States
Organisme : NCATS NIH HHS
ID : UG3 TR002881
Pays : United States
Organisme : NIGMS NIH HHS
ID : R35 GM119537
Pays : United States
Organisme : NHLBI NIH HHS
ID : UG3 HL147353
Pays : United States
Informations de copyright
Copyright © 2021 The Author(s). Published by Elsevier B.V. All rights reserved.
Références
Nucleic Acids Res. 2018 Jul 27;46(13):e80
pubmed: 29718399
Genome Biol. 2009;10(6):R64
pubmed: 19531210
Theranostics. 2017 Jan 15;7(3):634-646
pubmed: 28255356
Sci Rep. 2017 May 8;7(1):1550
pubmed: 28484218
Anal Bioanal Chem. 2016 Feb;408(5):1475-85
pubmed: 26700447
J Clin Invest. 2018 Jun 1;128(6):2551-2568
pubmed: 29757188
Sci Adv. 2019 Mar 13;5(3):eaau9443
pubmed: 30891499
Nature. 2005 Jun 16;435(7044):974-8
pubmed: 15944714
Chem Commun (Camb). 2009 Jun 7;(21):3065-7
pubmed: 19462088
Genet Anal. 1999 Feb;14(5-6):151-6
pubmed: 10084107
Nat Biotechnol. 2001 Apr;19(4):365-70
pubmed: 11283596
J Am Chem Soc. 2013 Sep 4;135(35):12952-5
pubmed: 23931073
Methods Mol Biol. 2017;1660:175-190
pubmed: 28828656
Proc Natl Acad Sci U S A. 1999 May 25;96(11):6171-6
pubmed: 10339560
Analyst. 2014 Jun 7;139(11):2867-72
pubmed: 24736939
iScience. 2020 Jan 24;23(1):100782
pubmed: 31958756
Nucleic Acids Res. 2019 Jan 8;47(D1):D155-D162
pubmed: 30423142
Nat Biotechnol. 1996 Mar;14(3):303-8
pubmed: 9630890
Curr Opin Chem Biol. 2004 Oct;8(5):547-53
pubmed: 15450499
Nat Methods. 2015 Feb;12(2):123-126
pubmed: 25486062
J Hepatol. 2017 May;66(5):1012-1021
pubmed: 28088579
Proc Natl Acad Sci U S A. 2018 Nov 13;115(46):E10849-E10858
pubmed: 30373812
IEEE Trans Nanobioscience. 2013 Sep;12(3):233-8
pubmed: 23722479
Proc Natl Acad Sci U S A. 2017 Sep 26;114(39):10367-10372
pubmed: 28893984
Cell. 1993 Dec 3;75(5):843-54
pubmed: 8252621
Biosens Bioelectron. 2011 Jan 15;26(5):2317-22
pubmed: 21115340
Nucleic Acids Res. 2005 Jan 19;33(2):e13
pubmed: 15659574
Mikrochim Acta. 2019 Feb 1;186(3):132
pubmed: 30707293
RNA. 2014 Aug;20(8):1183-94
pubmed: 24942625
Anal Chim Acta. 2019 Nov 4;1079:146-152
pubmed: 31387705
J Mater Chem B. 2017 Apr 28;5(16):3047-3058
pubmed: 32263996
Oncol Rep. 2016 Nov;36(5):3051-3057
pubmed: 27599551
Annu Rev Cell Dev Biol. 2007;23:175-205
pubmed: 17506695
Am J Cancer Res. 2016 Jul 01;6(7):1461-93
pubmed: 27508091
Appl Environ Microbiol. 2000 Aug;66(8):3603-7
pubmed: 10919826
Nucleic Acids Res. 2012 Jan;40(2):e13
pubmed: 22110035
Nucleic Acids Res. 2017 Jun 2;45(10):e87
pubmed: 28180294
Mol Pharm. 2012 Mar 5;9(3):685-93
pubmed: 22289057