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Total RNA sequencing in human biofluids and extracellular vesicles using SMARTer technology

ABSTRACT: Total RNA sequencing in human biofluids and extracellular vesicles using SMARTer technology

PROVIDER: PRJNA544444 | ENA |

REPOSITORIES: ENA

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Total RNA sequencing in human biofluids and extracellular vesicles using SMARTer technology

Project description:Transcriptome profiling has emerged as a powerful tool to investigate the biomarker potential of human biofluids. However, despite an enormous interest in extracellular nucleic acids, total RNA sequencing methods to quantify 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-poor plasma, urine, conditioned medium, and extracellular vesicles from these biofluids.

2019-09-01 | GSE131689 | GEO

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

Project description: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.

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Project description:SMARTer Stranded Total RNA-Seq method of human platelet-rich plasma, platelet-free plasma, urine, conditioned medium, and extracellular vesicles (EVs) from these biofluids. Including a titration experiment with spikes.

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Project description:Endogenous and exogenous extracellular RNAs will be characterized in two body fluids (plasma and CSF) collected at the same time from 180 healthy subjects ranging in age from 31 to 101 years old. Small RNAs contained in these samples will be profiled by small RNA-seq using unfractionated biofluids and purified extracellular vesicles.

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Large differences in small RNA composition between human biofluids - 10 biofluids

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