ArticleFrontiers in genetics2018
Modified Polyadenylation-Based RT-qPCR Increases Selectivity of Amplification of 3'-MicroRNA Isoforms.
Article in Frontiers in genetics, 2018. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 16 papers.
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Who cites it
16 citing papers in PubMed.
- IsomiR stoichiometry changes as disease biomarkers.Molecular therapy. Nucleic acids · 2025Article
- Review
- MBNL splicing factors regulate the microtranscriptome of skeletal muscles.Nucleic acids research · 2024Article
- 3'IsomiR Species Composition Affects Reliable Quantification of miRNA/isomiR Variants by Poly(A) RT-qPCR: Impact on Small RNA-Seq Profiling Validation.International journal of molecular sciences · 2023Article
- A comprehensive pan-cancer analysis reveals cancer-associated robust isomiR expression landscapes in miRNA arm switching.Molecular genetics and genomics : MGG · 2023Article
- MicroRNA 3' ends shorten during adolescent brain maturation.Frontiers in molecular neuroscience · 2023Article
- IsomiR-eQTL: A Cancer-Specific Expression Quantitative Trait Loci Database of miRNAs and Their Isoforms.International journal of molecular sciences · 2022Article
- Rapid and accurate quantification of isomiRs by RT-qPCR.Scientific reports · 2022Article
- Quantification of microRNA editing using two-tailed RT-qPCR for improved biomarker discovery.RNA (New York, N.Y.) · 2021Article
- mi-IsoNet: systems-scale microRNA landscape reveals rampant isoform-mediated gain of target interaction diversity and signaling specificity.Briefings in bioinformatics · 2021Article
- Genistein alleviates chronic vascular inflammatory response via the miR‑21/NF‑κB p65 axis in lipopolysaccharide‑treated mice.Molecular medicine reports · 2021Article
- Dynamic mRNP Remodeling in Response to Internal and External Stimuli.Biomolecules · 2020Review
- Stumbling on elusive cargo: how isomiRs challenge microRNA detection and quantification, the case of extracellular vesicles.Journal of extracellular vesicles · 2020Article
- miR-625-3p and lncRNA GAS5 in Liquid Biopsies for Predicting the Outcome of Malignant Pleural Mesothelioma Patients Treated with Neo-Adjuvant Chemotherapy and Surgery.Non-coding RNA · 2019Article
- An estimate of the total number of true human miRNAs.Nucleic acids research · 2019Article
- miRNA length variation during macrophage stimulation confounds the interpretation of results: implications for miRNA quantification by RT-qPCR.RNA (New York, N.Y.) · 2019Article
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4 authors.
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Abstract
MicroRNA (miRNA) detection by reverse transcription (RT) quantitative real-time PCR (RT-qPCR) is the most popular method currently used to measure miRNA expression. Although the majority of miRNA families are constituted of several 3'-end length variants ("isomiRs"), little attention has been paid to their differential detection by RT-qPCR. However, recent evidence indicates that 3'-end miRNA isoforms can exhibit 3'-length specific regulatory functions, underlining the need to develop strategies to differentiate 3'-isomiRs by RT-qPCR approaches. We demonstrate here that polyadenylation-based RT-qPCR strategies targeted to 20-21 nt isoforms amplify entire miRNA families, but that primers targeted to >22 nt isoforms were specific to >21 nt isoforms. Based on this observation, we developed a simple method to increase selectivity of polyadenylation-based RT-qPCR assays toward shorter isoforms, and demonstrate its capacity to help distinguish short RNAs from longer ones, using synthetic RNAs and biological samples with altered isomiR stoichiometry. Our approach can be adapted to many polyadenylation-based RT-qPCR technologies already exiting, providing a convenient way to distinguish long and short 3'-isomiRs.
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