ArticleNucleic acids research2016
Polysome shift assay for direct measurement of miRNA inhibition by anti-miRNA drugs.
Article in Nucleic acids research, 2016. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 13 papers.
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Who cites it
13 citing papers in PubMed, 18 citations in OpenAlex.
- Harnessing the power of miRNAs for precision diagnosis and treatment of male infertility.Naunyn-Schmiedeberg's archives of pharmacology · 2025Review
- Article
- miRNA-27a is essential for bone remodeling by modulating p62-mediated osteoclast signaling.eLife · 2023Article
- Effective silencing of miR-126 after ischemic stroke by means of intravenous α-tocopherol-conjugated heteroduplex oligonucleotide in mice.Scientific reports · 2021Article
- The Roles of MicroRNAs in Male Infertility.International journal of molecular sciences · 2021Review
- Identifying the Translatome of Mouse NEBD-Stage Oocytes via SSP-Profiling; A Novel Polysome Fractionation Method.International journal of molecular sciences · 2020Article
- Discovery and preclinical evaluation of anti-miR-17 oligonucleotide RGLS4326 for the treatment of polycystic kidney disease.Nature communications · 2019Article
- Highly efficient silencing of microRNA by heteroduplex oligonucleotides.Nucleic acids research · 2019Article
- Motor neuron-derived microRNAs cause astrocyte dysfunction in amyotrophic lateral sclerosis.Brain : a journal of neurology · 2018Article
- Simultaneous visualization of the subfemtomolar expression of microRNA and microRNA target gene using HILO microscopy.Chemical science · 2017Article
- Therapeutic microRNAs in polycystic kidney disease.Current opinion in nephrology and hypertension · 2017Review
- Potent Anti-seizure Effects of Locked Nucleic Acid Antagomirs Targeting miR-134 in Multiple Mouse and Rat Models of Epilepsy.Molecular therapy. Nucleic acids · 2017Article
- microRNA-17 family promotes polycystic kidney disease progression through modulation of mitochondrial metabolism.Nature communications · 2017Article
Corrections and comments
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Authors and funding
9 authors at 2 institutions in 1 country.
Funding
No grant is acknowledged in the PubMed record.
Abstract
Anti-miRNA (anti-miR) oligonucleotide drugs are being developed to inhibit overactive miRNAs linked to disease. To help facilitate the transition from concept to clinic, new research tools are required. Here we report a novel method--miRNA Polysome Shift Assay (miPSA)--for direct measurement of miRNA engagement by anti-miR, which is more robust than conventional pharmacodynamics using downstream target gene derepression. The method takes advantage of size differences between active and inhibited miRNA complexes. Active miRNAs bind target mRNAs in high molecular weight polysome complexes, while inhibited miRNAs are sterically blocked by anti-miRs from forming this interaction. These two states can be assessed by fractionating tissue or cell lysates using differential ultracentrifugation through sucrose gradients. Accordingly, anti-miR treatment causes a specific shift of cognate miRNA from heavy to light density fractions. The magnitude of this shift is dose-responsive and maintains a linear relationship with downstream target gene derepression while providing a substantially higher dynamic window for aiding drug discovery. In contrast, we found that the commonly used 'RT-interference' approach, which assumes that inhibited miRNA is undetectable by RT-qPCR, can yield unreliable results that poorly reflect the binding stoichiometry of anti-miR to miRNA. We also demonstrate that the miPSA has additional utility in assessing anti-miR cross-reactivity with miRNAs sharing similar seed sequences.
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