Loading of Extracellular Vesicles with Chemically Stabilized Hydrophobic siRNAs for the Treatment of Disease in the Central Nervous System
RNA Therapeutics Institute; Program in Molecular Medicine; Department of Medicine
Biochemistry, Biophysics, and Structural Biology | Neuroscience and Neurobiology
Efficient delivery of oligonucleotide therapeutics, i.e., siRNAs, to the central nervous system represents a significant barrier to their clinical advancement for the treatment of neurological disorders. Small, endogenous extracellular vesicles were shown to be able to transport lipids, proteins and RNA between cells, including neurons. This natural trafficking ability gives extracellular vesicles the potential to be used as delivery vehicles for oligonucleotides, i.e., siRNAs. However, robust and scalable methods for loading of extracellular vesicles with oligonucleotide cargo are lacking. We describe a detailed protocol for the loading of hydrophobically modified siRNAs into extracellular vesicles upon simple co-incubation. We detail methods of the workflow from purification of extracellular vesicles to data analysis. This method may advance extracellular vesicles-based therapies for the treatment of a broad range of neurological disorders.
Extracellular vesicles, hydrophobically modified siRNA, neurological disorder, RNA interference, therapy, UMCCTS funding
Rights and Permissions
Copyright: © 2017 The Authors; exclusive licensee Bio-protocol LLC.
DOI of Published Version
Bio Protoc. 2017 Jun 20;7(20). pii: e2338. doi: 10.21769/BioProtoc.2338. Link to article on publisher's site
Haraszti, Reka A.; Coles, Andrew H.; Aronin, Neil; Khvorova, Anastasia; and Didiot, Marie C., "Loading of Extracellular Vesicles with Chemically Stabilized Hydrophobic siRNAs for the Treatment of Disease in the Central Nervous System" (2017). Open Access Articles. 3170.