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Determination of primary microRNA processing in clinical samples by targeted pri-miR-sequencing

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Item Type:Article
Title:Determination of primary microRNA processing in clinical samples by targeted pri-miR-sequencing
Creators Name:Conrad, T. and Ntini, E. and Lang, B. and Cozzuto, L. and Andersen, J.B. and Marquardt, J.U. and Ponomarenko, J. and Tartaglia, G.G. and Vang Orom, U.A.
Abstract:MicroRNA expression is important for gene regulation and deregulated microRNA expression is often observed in disease such as cancer. The processing of primary microRNA transcripts is an important regulatory step in microRNA biogenesis. Due to low expression level and association with chromatin primary microRNAs are challenging to study in clinical samples where input material is limited. Here, we present a high-sensitivity targeted method to determine processing efficiency of several hundred primary microRNAs from total RNA that requires relatively few RNA sequencing reads. We validate the method using RNA from HeLa cells and show the applicability to clinical samples by analyzing RNA from normal liver and hepatocellular carcinoma. We identify 24 primary microRNAs with significant changes in processing efficiency from normal liver to hepatocellular carcinoma, among those the highly expressed miRNA-122 and miRNA-21, demonstrating that differential processing of primary microRNAs is occurring and could be involved in disease. With our method presented here we provide means to study pri-miRNA processing in disease from clinical samples.
Keywords:miRNA Biogenesis, Primary miRNA, RNA Sequencing, Liver, HCC, Clinical Samples, miRNA Processing, microRNA
Publisher:Cold Spring Harbor Laboratory Press
Page Range:1726-1730
Date:November 2020
Official Publication:https://doi.org/10.1261/rna.076240.120
PubMed:View item in PubMed

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