Preparation and isolation of siRNA-loaded extracellular vesicles

Pieter Vader; Imre Mäger; Yi Lee; Joel Z. Nordin; Samir E L Andaloussi; Matthew J A Wood

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

Preparation and isolation of siRNA-loaded extracellular vesicles

Pieter Vader^*
, Imre Mäger
, Yi Lee
, Joel Z. Nordin
, Samir E L Andaloussi
, Matthew J A Wood

^*Corresponding author for this work

Research output: Chapter in Book/Report/Conference proceeding › Chapter › Academic › peer-review

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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 RNAimediating 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 language	English
Title of host publication	Exosomes and Microvesicles
Subtitle of host publication	Methods and Protocols
Publisher	Humana Press Inc.
Pages	197-204
Number of pages	8
ISBN (Electronic)	978-1-4939-6728-5
ISBN (Print)	978-1-4939-6726-1
DOIs	https://doi.org/10.1007/978-1-4939-6728-5_14
Publication status	Published - 2017

Publication series

Name	Methods in Molecular Biology
Volume	1545
ISSN (Print)	1064-3745

Keywords

Drug delivery
Exosomes
Extracellular vesicles
Microvesicles
siRNA
Size-exclusion chromatography

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

14Final published version, 199 KBLicence: Taverne

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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 RNAimediating 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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author = "Pieter Vader and Imre M{\"a}ger and Yi Lee and Nordin, \{Joel Z.\} and Andaloussi, \{Samir E L\} and Wood, \{Matthew J A\}",

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T1 - Preparation and isolation of siRNA-loaded extracellular vesicles

AU - Vader, Pieter

AU - Mäger, Imre

AU - Lee, Yi

AU - Nordin, Joel Z.

AU - Andaloussi, Samir E L

AU - Wood, Matthew J A

N1 - Publisher Copyright: © Springer Science+Business Media LLC 2017.

PY - 2017

Y1 - 2017

N2 - 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 RNAimediating 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.

AB - 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 RNAimediating 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.

KW - Drug delivery

KW - Exosomes

KW - Extracellular vesicles

KW - Microvesicles

KW - siRNA

KW - Size-exclusion chromatography

UR - https://www.scopus.com/pages/publications/85006459584

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

M3 - Chapter

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SN - 978-1-4939-6726-1

T3 - Methods in Molecular Biology

SP - 197

EP - 204

BT - Exosomes and Microvesicles

PB - Humana Press Inc.

ER -

Preparation and isolation of siRNA-loaded extracellular vesicles

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