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

• Published in: PloS 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
• Language: English
• 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
• ISSN: 1932-6203; 1932-6203
• DOI: 10.1371/journal.pone.0136133

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".