PLX162463

GSE126049: Phospho-small RNA-seq reveals circulating, extracellular mRNA/lncRNAs as potential biomarkers in human plasma: Healthy Cohort [HEALTHYPLASMA]

• Organsim human
• Type RNASEQ
• Target gene
• Project ARCHS4

Extracellular RNAs (exRNAs) in blood and other biofluids have attracted great interest as potential biomarkers in liquid biopsy applications, as well as for their potential biological functions. Whereas it is well-established that extracellular microRNAs are present in human blood circulation, the degree to which messenger RNAs (mRNA) and long noncoding RNAs (lncRNA) are represented in plasma is less clear. Here we report that mRNA and lncRNA species are present as small fragments in plasma that are not detected by standard small RNA-seq methods, because they lack 5-phosphorylation or carry 3-phosphorylation. We developed a modified sequencing protocol (termed phospho-sRNA-seq) that incorporates upfront RNA treatment with T4 polynucleotide kinase (which also has 3 phosphatase activity) and compared it to a standard small RNA-seq protocol, using as input both a pool of synthetic RNAs with diverse 5 and 3 end chemistries, as well exRNA isolated from human blood plasma. Using a custom, high-stringency pipeline for data analysis we identified mRNA and lncRNA transcriptome fingerprints in plasma, including multiple tissue-specific gene sets. In this cohort of healthy individuals, we demonstrate the improved capture of mRNA and lncRNA fragments using the phospho-sRNA-seq protocol. We find they primarily arise from mRNA exons, and that abundant fragments are consistently detected in multiple individuals. SOURCE: Muneesh Tewari (mtewari@med.umich.edu) - 4029 BSRB University of Michigan