Performance assessment of total RNA sequencing of human biofluids and extracellular vesicles
Performance assessment of total RNA sequencing of human biofluids and extracellular vesicles
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DOI：
10.1038/s41598-019-53892-x
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发表时间：
2019-11-26
影响因子：
4.6
通讯作者：
Vandesompele, Jo
中科院分区：
文献类型：
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作者：
Everaert, Celine;Helsmoortel, Hetty;Vandesompele, Jo
RNA分析已成为研究人类生物流体生物标志物潜力的有力工具。然而，尽管对细胞外核酸有着巨大的兴趣，但用于定量细胞外总RNA含量的RNA测序方法却很少见。在这里，我们评估了SMARTer Stranded Total RNA-Seq方法在人富血小板血浆、无血小板血浆、尿液、条件培养基和来自这些生物流体的细胞外囊泡（EV）中的性能。我们发现该方法准确，精确，与低输入量兼容，并且能够量化几千个基因。我们挑选了不同种类的RNA分子，包括mRNA、lncRNA、circRNA、miscRNA和假基因。值得注意的是，读段分布和基因含量在生物流体之间存在显著差异。总之，我们是第一个表明SMARTer方法可用于无偏地解开广泛的生物流体及其细胞外囊泡的完整转录组。
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.