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Int J Nanomedicine. 2017 Apr 18;12:3153-3170. doi: 10.2147/IJN.S133430. eCollection 2017.

Pseudotyping exosomes for enhanced protein delivery in mammalian cells.

Author information

1
Department of Bioengineering, Santa Clara University, Santa Clara.
2
Crown College, University of California at Santa Cruz, Santa Cruz.
3
Department of Bioengineering, University of California at Berkeley, Berkeley, CA, USA.

Abstract

Exosomes are cell-derived nanovesicles that hold promise as living vehicles for intracellular delivery of therapeutics to mammalian cells. This potential, however, is undermined by the lack of effective methods to load exosomes with therapeutic proteins and to facilitate their uptake by target cells. Here, we demonstrate how a vesicular stomatitis virus glycoprotein (VSVG) can both load protein cargo onto exosomes and increase their delivery ability via a pseudotyping mechanism. By fusing a set of fluorescent and luminescent reporters with VSVG, we show the successful targeting and incorporation of VSVG fusions into exosomes by gene transfection and fluorescence tracking. We subsequently validate our system by live cell imaging of VSVG and its participation in endosomes/exosomes that are ultimately released from transfected HEK293 cells. We show that VSVG pseudotyping of exosomes does not affect the size or distributions of the exosomes, and both the full-length VSVG and the VSVG without the ectodomain are shown to integrate into the exosomal membrane, suggesting that the ectodomain is not required for protein loading. Finally, exosomes pseudotyped with full-length VSVG are internalized by multiple-recipient cell types to a greater degree compared to exosomes loaded with VSVG without the ectodomain, confirming a role of the ectodomain in cell tropism. In summary, our work introduces a new genetically encoded pseudotyping platform to load and enhance the intracellular delivery of therapeutic proteins via exosome-based vehicles to target cells.

KEYWORDS:

VSVG; exosome; nanotechnology; protein delivery

PMID:
28458537
PMCID:
PMC5402897
DOI:
10.2147/IJN.S133430
[Indexed for MEDLINE]
Free PMC Article

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