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Profiling of Extrac...
Profiling of Extracellular Small RNAs Highlights a Strong Bias towards Non-Vesicular Secretion
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- Sork, Helena (author)
- Karolinska Institutet
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Conceicao, Mariana (author)
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- Corso, Giulia (author)
- Karolinska Institutet
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- Nordin, Joel (author)
- Karolinska Institutet
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Lee, Yi Xin Fiona (author)
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Krjutskov, Kaarel (author)
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- Orzechowski Westholm, Jakub (author)
- Stockholms universitet,Science for Life Laboratory (SciLifeLab),Institutionen för biokemi och biofysik
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Vader, Pieter (author)
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Pauwels, Marie (author)
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Vandenbroucke, Roosmarijn E. (author)
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Wood, Matthew J. A. (author)
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- EL Andaloussi, Samir (author)
- Karolinska Institutet
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Mager, Imre (author)
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(creator_code:org_t)
- 2021-06-18
- 2021
- English.
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In: Cells. - : MDPI AG. - 2073-4409. ; 10:6
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https://doi.org/10.3...
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https://www.mdpi.com...
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Abstract
Subject headings
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- The extracellular environment consists of a plethora of molecules, including extracellular miRNA that can be secreted in association with extracellular vesicles (EVs) or soluble protein complexes (non-EVs). Yet, interest in therapeutic short RNA carriers lies mainly in EVs, the vehicles conveying the great majority of the biological activity. Here, by overexpressing miRNA and shRNA sequences in parent cells and using size exclusion liquid chromatography (SEC) to separate the secretome into EV and non-EV fractions, we saw that >98% of overexpressed miRNA was secreted within the non-EV fraction. Furthermore, small RNA sequencing studies of native miRNA transcripts revealed that although the abundance of miRNAs in EVs, non-EVs and parent cells correlated well (R-2 = 0.69-0.87), quantitatively an outstanding 96.2-99.9% of total miRNA was secreted in the non-EV fraction. Nevertheless, though EVs contained only a fraction of secreted miRNAs, these molecules were stable at 37 degrees C in a serum-containing environment, indicating that if sufficient miRNA loading is achieved, EVs can remain delivery-competent for a prolonged period of time. This study suggests that the passive endogenous EV loading strategy might be a relatively wasteful way of loading miRNA to EVs, and active miRNA loading approaches are needed for developing advanced EV miRNA therapies in the future.
Subject headings
- NATURVETENSKAP -- Biologi (hsv//swe)
- NATURAL SCIENCES -- Biological Sciences (hsv//eng)
Keyword
- extracellular vesicles
- small RNA
- SEC
- extracellular RNA
- miRNA
Publication and Content Type
- ref (subject category)
- art (subject category)
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Cells
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- By the author/editor
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Sork, Helena
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Conceicao, Maria ...
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Corso, Giulia
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Nordin, Joel
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Lee, Yi Xin Fion ...
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Krjutskov, Kaare ...
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show more...
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Orzechowski West ...
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Vader, Pieter
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Pauwels, Marie
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Vandenbroucke, R ...
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Wood, Matthew J. ...
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EL Andaloussi, S ...
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Mager, Imre
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- About the subject
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- NATURAL SCIENCES
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NATURAL SCIENCES
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and Biological Scien ...
- Articles in the publication
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Cells
- By the university
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Stockholm University
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Karolinska Institutet