Preparation and Isolation of siRNA-Loaded Extracellular Vesicles

  • Pieter VaderEmail author
  • Imre Mäger
  • Yi Lee
  • Joel Z. Nordin
  • Samir E. L. Andaloussi
  • Matthew J. A. Wood
Part of the Methods in Molecular Biology book series (MIMB, volume 1545)


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.

Key words

Extracellular vesicles Exosomes Microvesicles siRNA Size-exclusion chromatography Drug delivery 



P.V. was supported by a Rubicon Fellowship from the Netherlands Organisation for Scientific Research (NWO). I.M. is supported by a Postdoctoral MOBILITAS Fellowship of the Estonian Science Foundation. Y.L. is funded by the Agency of Science, Technology and Research (A*STAR), Singapore. SELA is supported by the Swedish Research Council (VR-MED and EuroNanoMedII) as well as the Swedish Society of Medical Research (SSMF).


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Copyright information

© Springer Science+Business Media LLC 2017

Authors and Affiliations

  • Pieter Vader
    • 1
    • 2
    Email author
  • Imre Mäger
    • 1
    • 3
  • Yi Lee
    • 1
  • Joel Z. Nordin
    • 4
  • Samir E. L. Andaloussi
    • 1
    • 4
  • Matthew J. A. Wood
    • 1
  1. 1.Department of Physiology, Anatomy and GeneticsUniversity of OxfordOxfordUK
  2. 2.Department of Clinical Chemistry and HaematologyUMC UtrechtUtrechtThe Netherlands
  3. 3.Institute of TechnologyUniversity of TartuTartuEstonia
  4. 4.Department of Laboratory MedicineKarolinska InstitutetHuddingeSweden

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