TY - JOUR
T1 - Detection of Extracellular Vesicle RNA Using Molecular Beacons
AU - Oliveira, Getulio Pereira de
AU - Zigon, Eric
AU - Rogers, Gaenna
AU - Davodian, Danny
AU - Lu, Shulin
AU - Jovanovic-Talisman, Tijana
AU - Jones, Jennifer
AU - Tigges, John
AU - Tyagi, Sanjay
AU - Ghiran, Ionita Calin
PY - 2020/1/24
Y1 - 2020/1/24
N2 - 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.
AB - 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.
KW - Biochemistry Methods
KW - Biomolecules
KW - Cell Biology
KW - Functional Aspects of Cell Biology
UR - http://www.scopus.com/inward/record.url?scp=85077912802&partnerID=8YFLogxK
U2 - 10.1016/j.isci.2019.100782
DO - 10.1016/j.isci.2019.100782
M3 - Journal article
C2 - 31958756
SN - 2589-0042
VL - 23
JO - iScience
JF - iScience
IS - 1
M1 - 100782
ER -