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Quantitative Analysis of Extracellular Vesicle Release Using Artificial MicroRNAs

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Cell-Secreted Vesicles

Part of the book series: Methods in Molecular Biology ((MIMB,volume 2668))

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Abstract

Despite the widely used concept of extracellular vesicle (EV)-mediated intercellular communication, we are still far from understanding what is the exact role of such nanosized vesicles in human physiology and disease. Thus, development of new methods and tools that enable the study of fundamental EV biology is valuable for advancing the field. Typically, EV production and release are monitored using approaches that rely on either antibody-based FACS assays or genetically encoded fluorescent proteins. We previously devised artificially barcoded exosomal microRNAs (bEXOmiRs) that were used as high-throughput reporters of EV release. In the first part of this protocol, basic steps and considerations for the design and cloning of bEXOmiRs are explained in detail. Next, analysis of bEXOmiR expression and abundance in cells and isolated EVs is described.

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Abbreviations

bEXOmiRs:

barcoded exosomal microRNAs

EV:

extracellular vesicle

miRNAs:

microRNAs

nt:

Nucleotides

RT:

Reverse transcription

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Authors and Affiliations

  1. Departament de Biomedicina, Unitat de Biologia Cel·lular, Facultat de Medicina i Ciències de la Salut, Centre de Recerca Biomèdica CELLEX, Institut d’Investigacions Biomèdiques August Pi i Sunyer, Universitat de Barcelona, Barcelona, Spain

    Albert Lu

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  1. Albert Lu
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Correspondence to Albert Lu .

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Editors and Affiliations

  1. GeneCellNano, Oulu University, Oulu, Finland

    Seppo Vainio

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© 2023 The Author(s), under exclusive license to Springer Science+Business Media, LLC, part of Springer Nature

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Lu, A. (2023). Quantitative Analysis of Extracellular Vesicle Release Using Artificial MicroRNAs. In: Vainio, S. (eds) Cell-Secreted Vesicles. Methods in Molecular Biology, vol 2668. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-3203-1_14

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  • DOI: https://doi.org/10.1007/978-1-0716-3203-1_14

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  • Publisher Name: Humana, New York, NY

  • Print ISBN: 978-1-0716-3202-4

  • Online ISBN: 978-1-0716-3203-1

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