Year: 2019
Type: article
Abstract: 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.
Source: iScience
Institutions: Beth Israel Deaconess Medical Center, Harvard University, Aalborg University, City Of Hope National Medical Center, City of Hope, Beckman Research Institute, National Cancer Institute, Center for Cancer Research, Rutgers, The State University of New Jersey, Rutgers New Jersey Medical School
Language: English
Subfield: Molecular Biology
Domain: Life Sciences
Open Access status: gold
Estimated APC paid: $3,000
Awards: G4256485472, Integrative, multi-parametric characterization of the EV surface protein and nucleic acid landscape by nano-flow and sorting cytometry, G6838740781, G6955268327, Background free amplified single-molecule FISH for in situ and flow cytometric applications, G9000219117, G9000292209, G9000303584
OpenAlex ID: W2995801297