Detection of Extracellular Vesicle RNA Using Molecular Beacons.
Biochemistry Methods
Biomolecules
Cell Biology
Functional Aspects of Cell Biology
Journal
iScience
ISSN: 2589-0042
Titre abrégé: iScience
Pays: United States
ID NLM: 101724038
Informations de publication
Date de publication:
24 Jan 2020
24 Jan 2020
Historique:
received:
06
03
2019
revised:
02
12
2019
accepted:
12
12
2019
pubmed:
21
1
2020
medline:
21
1
2020
entrez:
21
1
2020
Statut:
ppublish
Résumé
Extracellular vesicles (EVs) have recently emerged as intercellular conveyors of biological information and disease biomarkers. Identification and characterization of RNA species in single EVs are currently challenging. Molecular beacons (MBs) represent an attractive means for detecting specific RNA molecules. Coupling the MBs to cell-penetrating peptides (CPPs) provides a fast, effective, and membrane-type agnostic means to deliver MBs across the plasma membrane and into the cytosol. Here, we generated RBCs-derived EVs by complement activation and tested the ability of MBs coupled with CPP to detect miRNAs from RBC-EVs. Our results showed that RBC and RBC-EVs miRNA-451a can be detected using MB-CPP, and the respective fluorescence levels can be measured by nano-flow cytometry. MB-based detection of RNA via nano-flow cytometry creates a powerful new analytical framework in which a simple addition of a reagent allows profiling of specific RNA species present within certain EV subsets.
Identifiants
pubmed: 31958756
pii: S2589-0042(19)30527-9
doi: 10.1016/j.isci.2019.100782
pmc: PMC6992906
pii:
doi:
Types de publication
Journal Article
Langues
eng
Pagination
100782Subventions
Organisme : NCI NIH HHS
ID : R01 CA227291
Pays : United States
Organisme : NHLBI NIH HHS
ID : U01 HL126497
Pays : United States
Organisme : NCI NIH HHS
ID : R01 CA218500
Pays : United States
Organisme : NCATS NIH HHS
ID : UG3 TR002881
Pays : United States
Organisme : NHLBI NIH HHS
ID : UG3 HL147353
Pays : United States
Informations de copyright
Copyright © 2019 The Author(s). Published by Elsevier Inc. All rights reserved.
Déclaration de conflit d'intérêts
Declaration of Interests The authors declare no competing interests.
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