title: Preparation and Isolation of siRNA-Loaded Extracellular Vesicles.
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.
authors.affiliation: Department of Physiology, Anatomy and Genetics, University of Oxford, Oxford, UK. pvader@umcutrecht.nl.|Department of Clinical Chemistry and Haematology, UMC Utrecht, Heidelberglaan 1000, Utrecht, 3584, CX, The Netherlands. pvader@umcutrecht.nl.
authors.forename: Pieter
authors.initials: P
authors.lastname: Vader
authors.affiliation: Department of Physiology, Anatomy and Genetics, University of Oxford, Oxford, UK.|Institute of Technology, University of Tartu, Tartu, Estonia.
authors.forename: Imre
authors.initials: I
authors.lastname: Mäger
authors.affiliation: Department of Physiology, Anatomy and Genetics, University of Oxford, Oxford, UK.
authors.forename: Yi
authors.initials: Y
authors.lastname: Lee
authors.affiliation: Department of Laboratory Medicine, Karolinska Institutet, Huddinge, Sweden.
authors.forename: Joel Z
authors.initials: JZ
authors.lastname: Nordin
authors.affiliation: Department of Physiology, Anatomy and Genetics, University of Oxford, Oxford, UK.|Department of Laboratory Medicine, Karolinska Institutet, Huddinge, Sweden.
authors.forename: Samir E L
authors.initials: SE
authors.lastname: Andaloussi
authors.affiliation: Department of Physiology, Anatomy and Genetics, University of Oxford, Oxford, UK.
authors.forename: Matthew J A
authors.initials: MJ
authors.lastname: Wood
mesh_terms: D002458:Cell Fractionation / Q000379:methods; D002460:Cell Line; D000067128:Extracellular Vesicles / Q000378:metabolism*; D006801:Humans; D016133:Polymerase Chain Reaction; D034741:RNA, Small Interfering / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism*
chemical_list: D034741:RNA, Small Interfering
keywords: Drug delivery; Exosomes; Extracellular vesicles; Microvesicles; Size-exclusion chromatography; siRNA