Document Detail

Exosome-mediated delivery of siRNA in vitro and in vivo.
MedLine Citation:
PMID:  23154783     Owner:  NLM     Status:  MEDLINE    
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 ~3 weeks.
Samir El-Andaloussi; Yi Lee; Samira Lakhal-Littleton; Jinghuan Li; Yiqi Seow; Chris Gardiner; Lydia Alvarez-Erviti; Ian L Sargent; Matthew J A Wood
Related Documents :
23976853 - Synthesis of novel tetravalent galactosylated dtpa-dspe and study on hepatocyte-targeti...
506553 - The effect of nifurtimox on experimental infections with trypanosomatidae other than tr...
7319193 - Preferential action of liposome-entrapped 1-(2-chloroethyl)-3-(4-methylcyclohexyl)-1-ni...
Publication Detail:
Type:  Journal Article; Research Support, Non-U.S. Gov't     Date:  2012-11-15
Journal Detail:
Title:  Nature protocols     Volume:  7     ISSN:  1750-2799     ISO Abbreviation:  Nat Protoc     Publication Date:  2012 Dec 
Date Detail:
Created Date:  2012-11-16     Completed Date:  2013-04-16     Revised Date:  2014-03-19    
Medline Journal Info:
Nlm Unique ID:  101284307     Medline TA:  Nat Protoc     Country:  England    
Other Details:
Languages:  eng     Pagination:  2112-26     Citation Subset:  IM    
Export Citation:
APA/MLA Format     Download EndNote     Download BibTex
MeSH Terms
Brain / metabolism
Exosomes / genetics*
Gene Silencing*
Gene Transfer Techniques*
Genetic Vectors
RNA, Small Interfering / administration & dosage*,  genetics
Transfection / methods
Grant Support
F-1101//Parkinson's UK; G-1109//Parkinson's UK; G0900887//Medical Research Council; GR087730//Wellcome Trust; MR/J003360/1//Medical Research Council
Reg. No./Substance:
0/RNA, Small Interfering

From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine

Previous Document:  Isolation of mouse mesenchymal stem cells on the basis of expression of Sca-1 and PDGFR-?.
Next Document:  The use of phage display to generate conformation-sensor recombinant antibodies.