doi: 10.1007/978-1-0716-2341-1_16.
Affiliations
Item in Clipboard
Methods Mol Biol.
2022.
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.
Keywords:
Electroporation; Exosome; Extracellular; Loading; Protocol; Therapy; Transfection; Vesicle; miRNA; microRNA.
© 2022. The Author(s), under exclusive license to Springer Science+Business Media, LLC, part of Springer Nature.
References
-
-
Yáñez-Mó M, Siljander PR, Andreu Z et al (2015) Biological properties of extracellular vesicles and their physiological functions. J Extracell Vesicles 4:27066. doi.org/10.3402/jev.v4.27066
–
DOI
–
PubMed
-
-
-
Armstrong JP, Holme MN, Stevens MM (2017) Re-engineering extracellular vesicles as smart nanoscale therapeutics. ACS Nano 11(1):69–83. doi.org/10.1021/acsnano.6b07607
–
DOI
–
PubMed
–
PMC
-
-
-
Di Leva G, Garofalo M, Croce CM (2014) MicroRNAs in cancer. Annu Rev Pathol 9:287–314. doi.org/10.1146/annurev-pathol-012513-104715
–
DOI
–
PubMed
-
-
-
Romero-Cordoba SL, Salido-Guadarrama I, Rodriguez-Dorantes M, Hidalgo-Miranda A (2014) miRNA biogenesis: biological impact in the development of cancer. Cancer Biol Ther 15(11):1444–1455. doi.org/10.4161/15384047.2014.955442
–
DOI
–
PubMed
–
PMC
-
-
-
Qian H, Tay CY, Setyawati MI, Chia SL, Lee DS, Leong DT (2017) Protecting microRNAs from RNase degradation with steric DNA nanostructures. Chem Sci 8(2):1062–1067. doi.org/10.1039/c6sc01829g
–
DOI
–
PubMed
-
MeSH terms
Read more here: Source link