Evidence map›Paper›PMID 42215555›Full record

ArticleScientific reports2026

A combined PIE-FRET and FCS assay to monitor RNA dynamics and cleavage by SARS-CoV-2 Nsp15.

Kenya Gordon, Zoe M Wright, Cameron R Stephens, Meredith N Frazier, Benjamin S Clark, Isha M Wilson, Irene Silvernail, Robin E Stanley, Sharonda J LeBlanc

Abstract read
PubMed Full text at PMC Publisher
In one paragraph

Article in Scientific reports, 2026. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 1 paper.

0numbers the graph read from it
0cells of the map it votes in
1citing papers in PubMed
–field-weighted citation impact
1 · What the graph read from it

What it found

Each row is one number read from the abstract, on the scale the paper reported it, with its interval. Left of the dashed line favours the treatment, right favours the comparator. Under each row is the sentence it came from. New to these charts? A ten-minute tutorial.

The abstract states no effect estimate the extractor could read, or names no intervention and outcome on the map, so this paper lights no cell and moves no belief. It is still indexed, cited and linked below.

2 · The registry

The trial behind it

Trials whose registry record cites this paper, or whose number appears in the abstract. A trial that started after this paper was published is citing it as background, not reporting it.

Neither the registry nor the abstract names a trial number. If this is a trial report, that itself is worth knowing.

3 · Its place in the literature

Who cites it

1 citing paper in PubMed.

  1. Article
4 · The record

Corrections and comments

5 · Who and what money

Authors and funding

9 authors.

Kenya GordonDepartment of Physics and Astronomy, North Carolina State University, Raleigh, NC, 27695-8202, USA.
Zoe M WrightMolecular and Cellular Biology Laboratory, Department of Health and Human Services, National Institute of Environmental Health Sciences, National Institutes of Health, 111 T. W. Alexander Drive, Research Triangle Park, NC, 27709, USA.
Cameron R StephensDepartment of Physics and Astronomy, North Carolina State University, Raleigh, NC, 27695-8202, USA.
Meredith N FrazierMolecular and Cellular Biology Laboratory, Department of Health and Human Services, National Institute of Environmental Health Sciences, National Institutes of Health, 111 T. W. Alexander Drive, Research Triangle Park, NC, 27709, USA.
Benjamin S ClarkDepartment of Physics and Astronomy, North Carolina State University, Raleigh, NC, 27695-8202, USA.
Isha M WilsonMolecular and Cellular Biology Laboratory, Department of Health and Human Services, National Institute of Environmental Health Sciences, National Institutes of Health, 111 T. W. Alexander Drive, Research Triangle Park, NC, 27709, USA.
Irene SilvernailDepartment of Physics and Astronomy, North Carolina State University, Raleigh, NC, 27695-8202, USA.
Robin E StanleyMolecular and Cellular Biology Laboratory, Department of Health and Human Services, National Institute of Environmental Health Sciences, National Institutes of Health, 111 T. W. Alexander Drive, Research Triangle Park, NC, 27709, USA.
Sharonda J LeBlancDepartment of Physics and Astronomy, North Carolina State University, Raleigh, NC, 27695-8202, USA. sleblan@ncsu.edu.

Funding

Structural and Functional Characterization of the SARS-CoV-2 Endoribonuclease Nsp15ZIAES103340 · NIEHS · NATIONAL INSTITUTE OF ENVIRONMENTAL HEALTH SCIENCES · PI STANLEY, ROBIN · 2020 to 2025
$3.6M
Intramural NIH HHS ZIA ES103340NIEHS NIH HHS ZIA ES103340
6 · The paper itself

Abstract

Nsp15 is an endoribonuclease, highly conserved among coronaviruses, that helps the virus evade detection in host cells by cleaving uridine-rich viral RNA sequences. Those sequences would otherwise trigger immune response pathways. Its essential role in pathogenesis and highly conserved nature make Nsp15 an attractive target for therapeutic intervention. While its crystal structure and uridine specificity are well-established, the influence of RNA structural dynamics and divalent cations on Nsp15 activity remains less understood. Leveraging single-molecule Pulsed Interleaved Excitation (PIE)-FRET and Fluorescence Correlation Spectroscopy (FCS) in combination, we developed an assay to track RNA cleavage by Nsp15 variants in real time and monitor the conformational dynamics of hybrid RNA/DNA substrates. Using our methodology and analysis strategies, we obtained clear indicators of RNA cleavage with both PIE-FRET and FCS data analysis. Our assay also revealed signatures of unique dynamic behavior in uridine-containing RNA substrates, indicating that divalent cations enhance substrate flexibility, which is associated with a faster observed reaction rate of RNA cleavage by Nsp15 in the presence of Mn

Indexed as

Fluorescence Resonance Energy TransferRNA, ViralSARS-CoV-2Viral Nonstructural ProteinsCOVID-19HumansNucleic Acid ConformationSingle Molecule ImagingSpectrometry, FluorescenceUridineORF1ab polyprotein, SARS-CoV-2RNA, ViralUridineViral Nonstructural ProteinsFCSFRETNsp15RNA dynamicsSingle-molecule

Identifiers

PMID42215555
PMCPMC13454198

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Registered trials

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Read under generation 80e0d062 · epoch 390. Bibliography from PubMed, PubMed Central and OpenAlex; grants from NIH RePORTER; trial links from ClinicalTrials.gov; estimates, votes and beliefs from the OpenQuestion graph.