Delivery of Small Interfering RNAs to Cells via Exosomes

Methods Mol Biol. 2016:1364:105-25. doi: 10.1007/978-1-4939-3112-5_10.

Abstract

Exosomes are small membrane bound vesicles between 30 and 100 nm in diameter of endocytic origin that are secreted into the extracellular environment by many different cell types. Exosomes play a role in intercellular communication by transferring proteins, lipids, and RNAs to recipient cells.Exosomes from human cells could be used as vectors to provide cells with therapeutic RNAs. Here we describe how exogenous small interfering RNAs may successfully be introduced into various kinds of human exosomes using electroporation and subsequently delivered to recipient cells. Methods used to confirm the presence of siRNA inside exosomes and cells are presented, such as flow cytometry, confocal microscopy, and Northern blot.

Keywords: Electroporation; Exosome delivery; Exosome uptake; Exosomes; Extracellular vesicles; Plasma exosomes; Post-transcriptional gene silencing; siRNA delivery.

MeSH terms

  • Blood Buffy Coat / cytology
  • Blotting, Northern
  • Blotting, Western
  • Cell Line, Tumor
  • Cell Separation
  • Drug Delivery Systems / methods*
  • Electroporation / methods*
  • Exosomes / metabolism*
  • Flow Cytometry
  • Humans
  • Lymphocytes / cytology
  • Microscopy, Confocal
  • Monocytes / cytology
  • RNA, Small Interfering / metabolism*

Substances

  • RNA, Small Interfering