Exosome-mediated delivery of siRNA in vitro and in vivo

被引：477

作者：

El-Andaloussi, Samir ^{[1
,2
]}

Lee, Yi ^{[1
]}

Lakhal-Littleton, Samira ^{[1
]}

Li, Jinghuan ^{[1
]}

Seow, Yiqi ^{[3
]}

Gardiner, Chris ^{[4
]}

Alvarez-Erviti, Lydia ^{[5
]}

Sargent, Ian L. ^{[4
]}

Wood, Matthew J. A. ^{[1
]}

机构：

[1] Univ Oxford, Dept Physiol Anat & Genet, Oxford, England

[2] Karolinska Inst, Dept Lab Med, Huddinge, Sweden

[3] Agcy Sci Technol & Res, Mol Engn Lab, Singapore, Singapore

[4] John Radcliffe Hosp, Nuffield Dept Obstet & Gynaecol, Oxford OX3 9DU, England

[5] UCL, Inst Neurol, Univ Dept Clin Neurosci, London, England

来源：

NATURE PROTOCOLS | 2012年 / 7卷 / 12期

基金：

英国惠康基金;

关键词：

MULTIVESICULAR BODIES; DENDRITIC CELLS; MESSENGER-RNA; MICRORNAS; MECHANISM; DISEASE; BRAIN;

D O I：

10.1038/nprot.2012.131

中图分类号：

Q5 [生物化学];

学科分类号：

071010 ; 081704 ;

摘要：

The use of small interfering RNAs (siRNAs) to induce gene silencing has opened a new avenue in drug discovery. However, their therapeutic potential is hampered by inadequate tissue-specific delivery. Exosomes are promising tools for drug delivery across different biological barriers. Here we show how exosomes derived from cultured cells can be harnessed for delivery of siRNA in vitro and in vivo. This protocol first describes the generation of targeted exosomes through transfection of an expression vector, comprising an exosomal protein fused with a peptide ligand. Next, we explain how to purify and characterize exosomes from transfected cell supernatant. Next, we detail crucial steps for loading siRNA into exosomes. Finally, we outline how to use exosomes to efficiently deliver siRNA in vitro and in vivo in mouse brain. Examples of anticipated results in which exosome-mediated siRNA delivery is evaluated by functional assays and imaging are also provided. The entire protocol takes similar to 3 weeks.

引用

页码：2112 / 2126

页数：15

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