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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.

Original publication

DOI

10.1007/978-1-4939-6728-5_14

Type

Journal article

Journal

Methods in molecular biology (Clifton, N.J.)

Publication Date

01/2017

Volume

1545

Pages

197 - 204

Addresses

Department of Physiology, Anatomy and Genetics, University of Oxford, Oxford, UK. pvader@umcutrecht.nl.

Keywords

Cell Line, Humans, RNA, Small Interfering, Cell Fractionation, Polymerase Chain Reaction, Extracellular Vesicles