Rapid and accurate quantification of isomiRs by RT-qPCR

Currently, microRNAs (miRs) are annotated as a single defined sequence (canonical), even though high-throughput small RNA sequencing has identified miR isoforms (isomiRs) that differ from their canonical counterparts in length, sequence, or both. Here we describe a simple reverse transcriptase-quant...

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Bibliographic Details
Published in:Scientific reports 2022-10, Vol.12 (1), p.17220-17220, Article 17220
Main Authors: Franco, Sandra, Pluvinet, Raquel, Sanchez-Herrero, Jose Francisco, Sumoy, Lauro, Martinez, Miguel Angel
Format: Article
Language:English
Subjects:
631/1647
631/326
631/337
Adenosine
Gene expression
Hepatitis C
High-Throughput Nucleotide Sequencing
HIV
Human immunodeficiency virus
Humanities and Social Sciences
Humans
Isoforms
MicroRNAs
MicroRNAs - genetics
miRNA
multidisciplinary
Nucleotide sequence
Plasma
Polymerase chain reaction
Protein Isoforms - genetics
Real-Time Polymerase Chain Reaction
Ribonucleic acid
RNA
RNA-directed DNA polymerase
RNA-Directed DNA Polymerase - genetics
Science
Science (multidisciplinary)
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Summary:Currently, microRNAs (miRs) are annotated as a single defined sequence (canonical), even though high-throughput small RNA sequencing has identified miR isoforms (isomiRs) that differ from their canonical counterparts in length, sequence, or both. Here we describe a simple reverse transcriptase-quantitative polymerase chain reaction (RT-qPCR)-based assay for quantification of the miR-100-5p_iso_3p:−2 variant. We chose miR-100-5p because the canonical sequence was underrepresented in our evaluation of human plasma. The quantification of miR-100-5p_iso_3 p:−2 from 57 plasma samples demonstrated high concordance between high-throughput RNA sequencing and RT-qPCR results ( r  = 0.55, p  
ISSN:2045-2322
2045-2322
DOI:10.1038/s41598-022-22298-7