Restriction of SARS-CoV-2 Replication by Targeting Programmed −1 Ribosomal Frameshifting In Vitro
This article has been Reviewed by the following groups
Discuss this preprint
Start a discussion What are Sciety discussions?Listed in
- Evaluated articles (ScreenIT)
Abstract
Translation of open reading frame 1b (ORF1b) in severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) requires programmed −1 ribosomal frameshifting (−1 PRF) promoted by an RNA pseudoknot. The extent to which SARS-CoV-2 replication may be sensitive to changes in −1 PRF efficiency is currently unknown. Through an unbiased, reporter-based high-throughput compound screen, we identified merafloxacin, a fluoroquinolone antibacterial, as a −1 PRF inhibitor of SARS-CoV-2. Frameshift inhibition by merafloxacin is robust to mutations within the pseudoknot region and is similarly effective on −1 PRF of other beta coronaviruses. Importantly, frameshift inhibition by merafloxacin substantially impedes SARS-CoV-2 replication in Vero E6 cells, thereby providing the proof of principle of targeting −1 PRF as an effective antiviral strategy for SARS-CoV-2.
Article activity feed
-
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.
-
