Restriction of SARS-CoV-2 replication by targeting programmed −1 ribosomal frameshifting
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Abstract
A large variety of RNA viruses, including the novel coronavirus SARS-CoV-2, contain specific RNA structures that promote programmed ribosomal frameshifting (PRF) to regulate viral gene expression. From a high-throughput compound screen, we identified a PRF inhibitor for SARS-CoV-2 and found that it substantially impeded viral replication in cultured cells. Interestingly, the compound could target not only SARS-CoV-2 but also other coronaviruses that use similar RNA structures to promote frameshifting. These results suggest targeting PRF is a plausible, effective, and broad-spectrum antiviral strategy for SARS-CoV-2 and other coronaviruses.
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SciScore for 10.1101/2020.10.21.349225: (What is this?)
Please note, not all rigor criteria are appropriate for all manuscripts.
Table 1: Rigor
Institutional Review Board Statement not detected. Randomization not detected. Blinding not detected. Power Analysis not detected. Sex as a biological variable not detected. Cell Line Authentication not detected. Table 2: Resources
Antibodies Sentences Resources After 1-hour blocking with 5% nonfat dry milk in PBST, the membrane was incubated with primary antibodies (rabbit anti-C19ORF66, Invitrogen # PA5-59815; anti-C19ORF66suggested: (Thermo Fisher Scientific Cat# PA5-59815, RRID:AB_2638887); mouse anti-β-Actin) diluted (1:2000) in 5% milk/PBST at 4 °C with slow shaking overnight. anti-β-Actinsuggested: NoneExperimental Models: Cell Lines Sentences Resources Dual-luciferase PRF reporter assay: HeLa and HEK293T cells … SciScore for 10.1101/2020.10.21.349225: (What is this?)
Please note, not all rigor criteria are appropriate for all manuscripts.
Table 1: Rigor
Institutional Review Board Statement not detected. Randomization not detected. Blinding not detected. Power Analysis not detected. Sex as a biological variable not detected. Cell Line Authentication not detected. Table 2: Resources
Antibodies Sentences Resources After 1-hour blocking with 5% nonfat dry milk in PBST, the membrane was incubated with primary antibodies (rabbit anti-C19ORF66, Invitrogen # PA5-59815; anti-C19ORF66suggested: (Thermo Fisher Scientific Cat# PA5-59815, RRID:AB_2638887); mouse anti-β-Actin) diluted (1:2000) in 5% milk/PBST at 4 °C with slow shaking overnight. anti-β-Actinsuggested: NoneExperimental Models: Cell Lines Sentences Resources Dual-luciferase PRF reporter assay: HeLa and HEK293T cells were cultured in DMEM with 10% fetal bovine serum ( HeLasuggested: NoneHigh-throughput compound screen: HEK293T cells were plated in 384 well plates at the density of 5,000 cells/well. HEK293Tsuggested: NoneSARS-CoV-2 plaque formation assay: Vero E6 cells (ATCC) were seeded at 2 × 105 cells/well in 12-well plates. Vero E6suggested: NoneResults from OddPub: We did not detect open data. We also did not detect open code. Researchers are encouraged to share open data when possible (see Nature blog).
Results from LimitationRecognizer: An explicit section about the limitations of the techniques employed in this study was not found. We encourage authors to address study limitations.Results from TrialIdentifier: No clinical trial numbers were referenced.
Results from Barzooka: We did not find any issues relating to the usage of bar graphs.
Results from JetFighter: We did not find any issues relating to colormaps.
Results from rtransparent:- Thank you for including a conflict of interest statement. Authors are encouraged to include this statement when submitting to a journal.
- Thank you for including a funding statement. Authors are encouraged to include this statement when submitting to a journal.
- No protocol registration statement was detected.
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