电穿孔
细胞外小泡
小RNA
微泡
计算生物学
胞外囊泡
人口
生物
细胞生物学
纳米技术
材料科学
医学
基因
生物化学
环境卫生
作者
Margherita Alba Carlotta Pomatto,Federica Negro,Giovanni Camussi
出处
期刊:Methods in molecular biology
日期:2022-01-01
卷期号:: 219-230
被引量:3
标识
DOI:10.1007/978-1-0716-2341-1_16
摘要
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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