Characterization of RNA from Exosomes and Other Extracellular Vesicles Isolated by a Novel Spin Column-Based Method

Exosomes and other extracellular vesicles (commonly referred to as EVs) have generated a lot of attention for their potential applications in both diagnostics and therapeutics. The contents of these vesicles are the subject of intense research, and the relatively recent discovery of RNA inside EVs h...

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Published inPloS one Vol. 10; no. 8; p. e0136133
Main Authors Enderle, Daniel, Spiel, Alexandra, Coticchia, Christine M., Berghoff, Emily, Mueller, Romy, Schlumpberger, Martin, Sprenger-Haussels, Markus, Shaffer, Jonathan M., Lader, Eric, Skog, Johan, Noerholm, Mikkel
Format Journal Article
LanguageEnglish
Published United States Public Library of Science 28.08.2015
Public Library of Science (PLoS)
Subjects
Biomarkers
Breast cancer
Cancer
Cell-Derived Microparticles - chemistry
Cell-Derived Microparticles - metabolism
Deoxyribonucleic acid
DNA
Exosomes
Exosomes - chemistry
Exosomes - metabolism
Extracellular vesicles
Female
Gene expression
Humans
Lung cancer
Male
Metastasis
Mutation
Plasma
Prostate cancer
Proteomics
Reagent Kits, Diagnostic
Ribonucleic acid
RNA
RNA - blood
RNA - isolation & purification
Ultracentrifugation
Ultracentrifugation - methods
Vesicles
United States > US
Germany
Online AccessGet full text
ISSN1932-6203
1932-6203
DOI10.1371/journal.pone.0136133

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Summary:Exosomes and other extracellular vesicles (commonly referred to as EVs) have generated a lot of attention for their potential applications in both diagnostics and therapeutics. The contents of these vesicles are the subject of intense research, and the relatively recent discovery of RNA inside EVs has raised interest in the biological function of these RNAs as well as their potential as biomarkers for cancer and other diseases. Traditional ultracentrifugation-based protocols to isolate EVs are labor-intensive and subject to significant variability. Various attempts to develop methods with robust, reproducible performance have not yet been completely successful. Here, we report the development and characterization of a spin column-based method for the isolation of total RNA from EVs in serum and plasma. This method isolates highly pure RNA of equal or higher quantity compared to ultracentrifugation, with high specificity for vesicular over non-vesicular RNA. The spin columns have a capacity to handle up to 4 mL sample volume, enabling detection of low-abundance transcripts in serum and plasma. We conclude that the method is an improvement over traditional methods in providing a faster, more standardized way to achieve reliable high quality RNA preparations from EVs in biofluids such as serum and plasma. The first kit utilizing this new method has recently been made available by Qiagen as "exoRNeasy Serum/Plasma Maxi Kit".
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Competing Interests: All authors are full time employees of the for-profit organizations Exosome Diagnostics or QIAGEN. Exosome Diagnostics and QIAGEN provided support in the form of salaries for authors [DE, AS, CC, EB, RM, MS, MSH, JMS, EL, JS, MN], but did not have any additional role in the study design, data collection and analysis, decision to publish, or preparation of the manuscript. The specific roles of these authors are articulated in the ‘author contributions’ section. The authors DE, EB, JS and MN are listed as inventors of a patent application (Methods for isolating microvesicles, PCT/US2014/010173) relating to this work. This does not alter the authors' adherence to PLOS ONE policies on sharing data and materials.
Conceived and designed the experiments: DE CC MS JS MN. Performed the experiments: AS CC EB RM JMS. Analyzed the data: DE CC MS MSH EL JS MN. Wrote the paper: DE JS MN.
ISSN:1932-6203
1932-6203
DOI:10.1371/journal.pone.0136133