Your browser doesn't support javascript.
loading
Analyzing the miRNA content of extracellular vesicles by fluorescence nanoparticle tracking.
Baldwin, Scott; Deighan, Clayton; Bandeira, Elga; Kwak, Kwang J; Rahman, Mohammad; Nana-Sinkam, Patrick; Lee, L James; Paulaitis, Michael E.
Affiliation
  • Baldwin S; William G. Lowrie Department of Chemical and Biomolecular Engineering, Ohio State University, Columbus, OH, USA; Nanoscale Science and Engineering Center for Affordable Nanoengineering of Polymeric Biomedical Devices, Ohio State University, Columbus, OH, USA.
  • Deighan C; William G. Lowrie Department of Chemical and Biomolecular Engineering, Ohio State University, Columbus, OH, USA; Malvern Instruments, Westborough, MA, USA.
  • Bandeira E; Center for Nanomedicine, Johns Hopkins University School of Medicine, Baltimore, MD, USA; Institute of Biophysics Carlos Chagas Filho, Federal University of Rio de Janeiro, Rio de Janeiro, Brazil.
  • Kwak KJ; William G. Lowrie Department of Chemical and Biomolecular Engineering, Ohio State University, Columbus, OH, USA; Nanoscale Science and Engineering Center for Affordable Nanoengineering of Polymeric Biomedical Devices, Ohio State University, Columbus, OH, USA.
  • Rahman M; Division of Pulmonary, Allergy, Critical Care and Sleep Medicine, Wexner Medical Center, Ohio State University, Columbus, OH, USA.
  • Nana-Sinkam P; Division of Pulmonary, Allergy, Critical Care and Sleep Medicine, Wexner Medical Center, Ohio State University, Columbus, OH, USA.
  • Lee LJ; William G. Lowrie Department of Chemical and Biomolecular Engineering, Ohio State University, Columbus, OH, USA; Nanoscale Science and Engineering Center for Affordable Nanoengineering of Polymeric Biomedical Devices, Ohio State University, Columbus, OH, USA.
  • Paulaitis ME; William G. Lowrie Department of Chemical and Biomolecular Engineering, Ohio State University, Columbus, OH, USA; Nanoscale Science and Engineering Center for Affordable Nanoengineering of Polymeric Biomedical Devices, Ohio State University, Columbus, OH, USA; Center for Nanomedicine, Johns Hopkins U
Nanomedicine ; 13(2): 765-770, 2017 02.
Article in En | MEDLINE | ID: mdl-27989853
We present a method that takes advantage of the fluorophore loading dependence of fluorescence nanoparticle tracking (fNTA) to determine the content of specific miRNA targets in extracellular vesicles (EVs) and their stoichiometry across the entire EV population. The method is based on an assay for detecting EV miRNA by hybridization to fluorescently labeled, miRNA-specific molecular beacons encapsulated in cationic lipoplex nanoparticles that fuse non-specifically with negatively charged EVs. To demonstrate the method, we carry out a stoichiometric analysis of miR-21 in EVs released from A549 lung cancer cells. We find approximately 2.3% of the A549 EVs have an average copy number of ~44 miR-21/A549 EV and contain at least a threshold number of 33 miR-21 copies/A549 EV required for fluorescence tracking. Potential applications of sizing, enumerating, and phenotyping EVs using this method include specifying dosages for therapeutic applications and identifying specific EV subpopulations in patient samples for diagnostic applications.
Subject(s)
Key words
Fulltext
Add to My VHL
Print
XML
PubMed Links
Search on Google

Full text: 1 Database: MEDLINE Main subject: MicroRNAs / Nanoparticles / Fluorescence Limits: Humans Language: En Journal: Nanomedicine Journal subject: BIOTECNOLOGIA Year: 2017 Type: Article Affiliation country: United States
Fulltext
Add to My VHL
Print
XML
PubMed Links
Search on Google

Full text: 1 Database: MEDLINE Main subject: MicroRNAs / Nanoparticles / Fluorescence Limits: Humans Language: En Journal: Nanomedicine Journal subject: BIOTECNOLOGIA Year: 2017 Type: Article Affiliation country: United States