Preparation and Isolation of siRNA-Loaded Extracellular Vesicles

Methods Mol Biol. 2017:1545:197-204. doi: 10.1007/978-1-4939-6728-5_14.

Abstract

RNA interference (RNAi) has tremendous potential for specific silencing of disease-causing genes. Its clinical usage however critically depends on the development of carrier systems that can transport the RNAi-mediating small interfering RNA (siRNA) molecules to the cytosol of target cells. Recent reports have suggested that extracellular vesicles (EVs) form a natural transport system through which biomolecules, including RNA, is exchanged between cells. Therefore, EVs are increasingly being considered as potential therapeutic siRNA delivery systems.In this chapter we describe a method for preparing siRNA-loaded EVs, including a robust, scalable method to isolate them from cell culture supernatants.

Keywords: Drug delivery; Exosomes; Extracellular vesicles; Microvesicles; Size-exclusion chromatography; siRNA.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Cell Fractionation* / methods
  • Cell Line
  • Extracellular Vesicles / metabolism*
  • Humans
  • Polymerase Chain Reaction
  • RNA, Small Interfering / genetics
  • RNA, Small Interfering / isolation & purification
  • RNA, Small Interfering / metabolism*

Substances

  • RNA, Small Interfering