Preparation and Isolation of siRNA-Loaded Extracellular Vesicles
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 wordsExtracellular 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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