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Determination of primary microRNA processing in clinical samples by targeted pri-miR-sequencing
- Source :
- Conrad, T, Ntini, E, Lang, B, Cozzuto, L, Andersen, J B, Marquardt, J U, Ponomarenko, J, Tartaglia, G G & Vang Orom, U A 2020, ' Determination of primary microRNA processing in clinical samples by targeted pri-miR-sequencing ', R N A, vol. 26, no. 11, pp. 1726-1730 . https://doi.org/10.1261/rna.076240.120, Conrad, T, Ntini, E, Lang, B, Cozzuto, L, Andersen, J B, Marquardt, J U, Ponomarenko, J, Tartaglia, G G & Ørom, U A V 2020, ' Determination of primary microRNA processing in clinical samples by targeted pri-miR-sequencing ', RNA, vol. 26, no. 11, pp. 1726-1730 . https://doi.org/10.1261/rna.076240.120, RNA
- Publication Year :
- 2020
- Publisher :
- Cold Spring Harbor Laboratory, 2020.
-
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 using as little as 500 thousand Illumina HiSeq 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.
- Subjects :
- Carcinoma, Hepatocellular
Clinical samples
Method
Computational biology
Disease
Biology
HCC
RNA sequencing
clinical samples
liver
miRNA biogenesis
primary miRNAs
HeLa
03 medical and health sciences
microRNA
medicine
Humans
Primary miRNAs
Molecular Biology
030304 developmental biology
Regulation of gene expression
0303 health sciences
Primary MicroRNA
Sequence Analysis, RNA
Liver Neoplasms
030302 biochemistry & molecular biology
High-Throughput Nucleotide Sequencing
Cancer
RNA
medicine.disease
biology.organism_classification
Chromatin
Gene Expression Regulation, Neoplastic
MicroRNAs
Liver
Hepatocellular carcinoma
HeLa Cells
Subjects
Details
- ISSN :
- 14699001 and 13558382
- Volume :
- 26
- Database :
- OpenAIRE
- Journal :
- RNA
- Accession number :
- edsair.doi.dedup.....408556077aa6db09a0395a8081fd2f61