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Preparation and Isolation of siRNA-Loaded Extracellular Vesicles

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Exosomes and Microvesicles

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1545))

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.

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Acknowledgments

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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Authors and Affiliations

  1. Department of Physiology, Anatomy and Genetics, University of Oxford, Oxford, UK

    Pieter Vader, Imre Mäger, Yi Lee, Samir E. L. Andaloussi & Matthew J. A. Wood

  2. Department of Clinical Chemistry and Haematology, UMC Utrecht, Heidelberglaan 1000, Utrecht, 3584, CX, The Netherlands

    Pieter Vader

  3. Institute of Technology, University of Tartu, Tartu, Estonia

    Imre Mäger

  4. Department of Laboratory Medicine, Karolinska Institutet, Huddinge, Sweden

    Joel Z. Nordin & Samir E. L. Andaloussi

Authors
  1. Pieter Vader
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  2. Imre Mäger
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  3. Yi Lee
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  4. Joel Z. Nordin
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  5. Samir E. L. Andaloussi
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  6. Matthew J. A. Wood
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Corresponding author

Correspondence to Pieter Vader .

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Editors and Affiliations

  1. Department of Biochemistry and Genetics, La Trobe Institute for Molecular Science, La Trobe University, Bundoora, Victoria, Australia

    Andrew F Hill

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Vader, P., Mäger, I., Lee, Y., Nordin, J.Z., Andaloussi, S.E.L., Wood, M.J.A. (2017). Preparation and Isolation of siRNA-Loaded Extracellular Vesicles. In: Hill, A. (eds) Exosomes and Microvesicles. Methods in Molecular Biology, vol 1545. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-6728-5_14

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  • DOI: https://doi.org/10.1007/978-1-4939-6728-5_14

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  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-6726-1

  • Online ISBN: 978-1-4939-6728-5

  • eBook Packages: Springer Protocols

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