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Performance assessment of total RNA sequencing of human biofluids and extracellular vesicles

Celine Everaert (UGent) , Hetty Helsmoortel (UGent) , Anneleen Decock (UGent) , Eva Hulstaert (UGent) , Ruben Van Paemel (UGent) , Kimberly Verniers (UGent) , Justine Nuytens (UGent) , Jasper Anckaert (UGent) , Nele Nijs (UGent) , Joeri Tulkens (UGent) , et al.
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Abstract
RNA profiling has emerged as a powerful tool to investigate the biomarker potential of human biofluids. However, despite enormous interest in extracellular nucleic acids, RNA sequencing methods to quantify the total RNA content outside cells are rare. Here, we evaluate the performance of the SMARTer Stranded Total RNA-Seq method in human platelet-rich plasma, platelet-free plasma, urine, conditioned medium, and extracellular vesicles (EVs) from these biofluids. We found the method to be accurate, precise, compatible with low-input volumes and able to quantify a few thousand genes. We picked up distinct classes of RNA molecules, including mRNA, lncRNA, circRNA, miscRNA and pseudogenes. Notably, the read distribution and gene content drastically differ among biofluids. In conclusion, we are the first to show that the SMARTer method can be used for unbiased unraveling of the complete transcriptome of a wide range of biofluids and their extracellular vesicles.
Keywords
LIQUID BIOPSIES, LANDSCAPE

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MLA
Everaert, Celine, et al. “Performance Assessment of Total RNA Sequencing of Human Biofluids and Extracellular Vesicles.” SCIENTIFIC REPORTS, vol. 9, 2019.
APA
Everaert, C., Helsmoortel, H., Decock, A., Hulstaert, E., Van Paemel, R., Verniers, K., … Vandesompele, J. (2019). Performance assessment of total RNA sequencing of human biofluids and extracellular vesicles. SCIENTIFIC REPORTS, 9.
Chicago author-date
Everaert, Celine, Hetty Helsmoortel, Anneleen Decock, Eva Hulstaert, Ruben Van Paemel, Kimberly Verniers, Justine Nuytens, et al. 2019. “Performance Assessment of Total RNA Sequencing of Human Biofluids and Extracellular Vesicles.” SCIENTIFIC REPORTS 9.
Chicago author-date (all authors)
Everaert, Celine, Hetty Helsmoortel, Anneleen Decock, Eva Hulstaert, Ruben Van Paemel, Kimberly Verniers, Justine Nuytens, Jasper Anckaert, Nele Nijs, Joeri Tulkens, Bert Dhondt, An Hendrix, Pieter Mestdagh, and Jo Vandesompele. 2019. “Performance Assessment of Total RNA Sequencing of Human Biofluids and Extracellular Vesicles.” SCIENTIFIC REPORTS 9.
Vancouver
1.
Everaert C, Helsmoortel H, Decock A, Hulstaert E, Van Paemel R, Verniers K, et al. Performance assessment of total RNA sequencing of human biofluids and extracellular vesicles. SCIENTIFIC REPORTS. 2019;9.
IEEE
[1]
C. Everaert et al., “Performance assessment of total RNA sequencing of human biofluids and extracellular vesicles,” SCIENTIFIC REPORTS, vol. 9, 2019.
@article{8636967,
  abstract     = {RNA profiling has emerged as a powerful tool to investigate the biomarker potential of human biofluids. However, despite enormous interest in extracellular nucleic acids, RNA sequencing methods to quantify the total RNA content outside cells are rare. Here, we evaluate the performance of the SMARTer Stranded Total RNA-Seq method in human platelet-rich plasma, platelet-free plasma, urine, conditioned medium, and extracellular vesicles (EVs) from these biofluids. We found the method to be accurate, precise, compatible with low-input volumes and able to quantify a few thousand genes. We picked up distinct classes of RNA molecules, including mRNA, lncRNA, circRNA, miscRNA and pseudogenes. Notably, the read distribution and gene content drastically differ among biofluids. In conclusion, we are the first to show that the SMARTer method can be used for unbiased unraveling of the complete transcriptome of a wide range of biofluids and their extracellular vesicles.},
  articleno    = {17574},
  author       = {Everaert, Celine and Helsmoortel, Hetty and Decock, Anneleen and Hulstaert, Eva and Van Paemel, Ruben and Verniers, Kimberly and Nuytens, Justine and Anckaert, Jasper and Nijs, Nele and Tulkens, Joeri and Dhondt, Bert and Hendrix, An and Mestdagh, Pieter and Vandesompele, Jo},
  issn         = {2045-2322},
  journal      = {SCIENTIFIC REPORTS},
  keywords     = {LIQUID BIOPSIES,LANDSCAPE},
  language     = {eng},
  pages        = {16},
  title        = {Performance assessment of total RNA sequencing of human biofluids and extracellular vesicles},
  url          = {http://dx.doi.org/10.1038/s41598-019-53892-x},
  volume       = {9},
  year         = {2019},
}

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