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Title: Exogenous loading of miRNAs into small extracellular vesicles
Authors: Abreu, Ricardo C de 
Ramos, Cristiana V
Becher, Clarissa
Lino, Miguel
Jesus, Carlos
Martins, Paula A da Costa
Martins, Patrícia A T
Moreno, Maria João 
Fernandes, Hugo
Ferreira, Lino
Keywords: extracellular vesicles; microRNA; modulation; post‐isolation
Issue Date: Aug-2021
Project: 007630 UID/QUI/00313/2019 
Programa Operacional Regional do Centro’ CENTRO2020 
Projects Interreg entitled: ‘Impulso de una red de I+i en química biológica para diagnóstico y tratamiento de enfermedades neurológicas 
EAPA_791/2018 - NEUROATLANTIC entitled: ‘An Atlantic innovation platform on diagnosis and treatment of neurological diseases and aging’ 
info:eu-repo/grantAgreement/EC/H2020/952266/EU/RESEarch for healThy AGEING 
Serial title, monograph or event: J Extracell Vesicles
Volume: 10
Issue: 10
Abstract: Small extracellular vesicles (sEVs), through their natural ability to interact with biological membranes and exploit endogenous processing pathways to convey biological information, are quintessential for the delivery of therapeutically relevant compounds, such as microRNAs (miRNAs) and proteins. Here, we used a fluorescently-labelled miRNA to quantify the efficiency of different methods to modulate the cargo of sEVs. Our results showed that, compared with electroporation, heat shock, permeation by a detergent-based compound (saponin) or cholesterol-modification of the miRNA, Exo-Fect was the most efficient method with > 50% transfection efficiency. Furthermore, qRT-PCR data showed that, compared with native sEVs, Exo-Fect modulation led to a > 1000-fold upregulation of the miRNA of interest. Importantly, this upregulation was observed for sEVs isolated from multiple sources. The modulated sEVs were able to delivery miR-155-5p into a reporter cell line, confirming the successful delivery of the miRNA to the target cell and, more importantly, its functionality. Finally, we showed that the membrane of Exo-Fect-loaded sEVs was altered compared with native sEVs and that enhanced the internalization of Exo-Fect-loaded sEVs within the target cells and decreased the interaction of those modulated sEVs with lysosomes.
ISSN: 2001-3078
DOI: 10.1002/jev2.12111
Rights: openAccess
Appears in Collections:UC Bibliotecas - Artigos em Revistas Internacionais

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