@article{0e9a05fa383c46b8b54fce7d7c4a6527,
title = "Exogenous loading of miRNAs into small extracellular vesicles",
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.",
keywords = "DELIVERY, ELECTROPORATION, ENDOSOMES, EXOSOMES, EXPRESSION, FLUORESCENCE, INHIBITOR, NANOPARTICLES, SIRNA, STEM-CELLS, extracellular vesicles, microRNA, modulation, post-isolation",
author = "Abreu, {Ricardo C.} and Cristiana Ramos and Clarissa Becher and Miguel Lino and Carlos Jesus and {da Costa Martins}, {Paula A.} and Martins, {Patricia A. T.} and Moreno, {Maria Joao} and Hugo Fernandes and Lino Ferreira",
note = "Funding Information: This work was partially supported by the Portuguese {\textquoteleft}Funda{\c c}{\~a}o para a Ci{\^e}ncia e a Tecnologia{\textquoteright} (FCT) through projects 007630 UID/QUI/00313/2019, PT2020_PTDC_DTP-FTO_2784_2014, POCI-01-0145-FEDER-029919, co-funded by COMPETE2020-UE and CENTRO-01-0145-FEDER-000014 through {\textquoteleft}Programa Operacional Regional do Centro{\textquoteright} CENTRO2020; Projects Interreg entitled: {\textquoteleft}Impulso de una red de I+i en qu{\'i}mica biol{\'o}gica para diagn{\'o}stico y tratamiento de enfermedades neurol{\'o}gicas{\textquoteright} and EAPA_791/2018 - NEUROATLANTIC entitled: {\textquoteleft}An Atlantic innovation platform on diagnosis and treatment of neurological diseases and aging{\textquoteright} and European Union{\textquoteright}s Horizon 2020 research and innovation program under grant agreement No 952266 (Project RESETageing). RA was supported by FCT (SFRH/BD/129317/2017). We would like to acknowledge Crioestaminal (www.crioestaminal.pt) for MSC samples. Funding Information: This work was partially supported by the Portuguese {\textquoteleft}Funda{\c c}{\~a}o para a Ci{\^e}ncia e a Tecnologia{\textquoteright} (FCT) through projects 007630 UID/QUI/00313/2019, PT2020_PTDC_DTP‐FTO_2784_2014, POCI‐01‐0145‐FEDER‐029919, co‐funded by COMPETE2020‐UE and CENTRO‐01‐0145‐FEDER‐000014 through {\textquoteleft}Programa Operacional Regional do Centro{\textquoteright} CENTRO2020; Projects Interreg entitled: {\textquoteleft}Impulso de una red de I+i en qu{\'i}mica biol{\'o}gica para diagn{\'o}stico y tratamiento de enfermedades neurol{\'o}gicas{\textquoteright} and EAPA_791/2018 ‐ NEUROATLANTIC entitled: {\textquoteleft}An Atlantic innovation platform on diagnosis and treatment of neurological diseases and aging{\textquoteright} and European Union{\textquoteright}s Horizon 2020 research and innovation program under grant agreement No 952266 (Project RESETageing). RA was supported by FCT (SFRH/BD/129317/2017). We would like to acknowledge Crioestaminal ( www.crioestaminal.pt ) for MSC samples. Publisher Copyright: {\textcopyright} 2021 The Authors. Journal of Extracellular Vesicles published by Wiley Periodicals, LLC on behalf of the International Society for Extracellular Vesicles",
year = "2021",
month = aug,
doi = "10.1002/jev2.12111",
language = "English",
volume = "10",
journal = "Journal of Extracellular Vesicles",
issn = "2001-3078",
publisher = "Routledge/Taylor & Francis Group",
number = "10",
}