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Cited by 16Extracellular vesicles in diseaseRNA Interference and Gene DeliveryNanoparticle-Based Drug Delivery

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Optimized Protocol for Plasma-Derived Extracellular Vesicles Loading with Synthetic miRNA Mimic Using Electroporation

Pomatto, Margherita A. C., Negro, Federica, Camussi, Giovanni·2022·Methods in Molecular Biology
DOI 10.1007/978-1-0716-2341-1_16
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Abstract

Extracellular vesicles (EVs) are a population of particles naturally released by cells to transport biological messages, including nucleic acids. Thus, EVs represent an ideal vehicle to deliver therapeutic miRNAs. The current challenge is the development of efficient protocols to load EVs with exogenous miRNAs. Human plasma is an abundant source of EVs which can be manipulated for therapeutic applications. Despite numerous techniques are currently available to load EVs, all of them present issues which limit their clinical application. Among all, electroporation was shown to be superior to other protocols and to efficiently load plasma-derived EVs with miRNAs. However, also the electroporation procedure presents issues that can reduce the miRNA delivery. In this chapter, we describe a protocol to isolate EVs from human plasma, to load synthetic miRNA mimics using electroporation, to evaluate EV integrity and miRNA loading into EVs. Finally, the analysis of EV functionality allows to investigate the ability of engineered EVs to transfer the miRNAs to target cells and to exploit their biological effects.

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Research works printed but not linked: 19

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Metadata

Authors
Pomatto, M. A. C., Negro, F., & Camussi, G.
Affiliations
University of Turin (IT)
Corresponding author
Pomatto, Margherita A. C.
ORCID
Venue
Methods in Molecular Biology
Publisher
Springer US
Published
2022
Volume
2504
Pages
219-230
ISSN
  • 1064-3745 (ISSN-L)
  • 1940-6029
DOI
10.1007/978-1-0716-2341-1_16
OpenAlex
W4226149320
Cited by
16
References
0
Citations by year
2026: 4 · 2025: 4 · 2024: 3 · 2023: 4 · 2022: 1
FWCI
5.03
Type
book chapter
Language
en
Open access
closed
Licence
Locations
  • Methods in molecular biology (publishedVersion)
  • PubMed (publishedVersion)
Topics
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Extracellular vesicles in disease
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › RNA Interference and Gene Delivery
  • Physical Sciences › Materials Science › Biomaterials › Nanoparticle-Based Drug Delivery
Keywords
extracellular vesicles, plasma-derived vesicles, electroporation, microRNA delivery, vesicle loading, vesicle isolation, vesicle integrity, microRNA transfer
MeSH terms
Extracellular Vesicles, Humans, Nucleic Acids, Electroporation, MicroRNAs

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