Development of a Fluorescence-Based, High-Throughput SARS-CoV-2 3CL ^pro Reporter Assay

Article activity feed

1. Version published to 10.1128/jvi.01265-20

   Oct 27, 2020
2. 

   ScreenIT

   Jun 25, 2020

   SciScore for 10.1101/2020.06.24.169565: (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

   Experimental Models: Cell Lines
   Sentences	Resources
   Cell culture: All cells were obtained from ATCC and grown at 37°C in 5% CO2. 293T cells were grown in Dulbecco’s Modified Eagle Medium (DMEM) supplemented with 5% fetal bovine serum, GlutaMAX, and penicillin-streptomycin.	293T suggested: NCBI_Iran Cat# C498, RRID:CVCL_0063)
   Cytotoxicity Assays: 24 hours before treatment, plates were poly-lysine treated and seeded with 293T or VeroE6 cells.	VeroE6 suggested: JCRB Cat# JCRB1819, RRID:CVCL_YQ4…

   SciScore for 10.1101/2020.06.24.169565: (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

   Experimental Models: Cell Lines
   Sentences	Resources
   Cell culture: All cells were obtained from ATCC and grown at 37°C in 5% CO2. 293T cells were grown in Dulbecco’s Modified Eagle Medium (DMEM) supplemented with 5% fetal bovine serum, GlutaMAX, and penicillin-streptomycin.	293T suggested: NCBI_Iran Cat# C498, RRID:CVCL_0063)
   Cytotoxicity Assays: 24 hours before treatment, plates were poly-lysine treated and seeded with 293T or VeroE6 cells.	VeroE6 suggested: JCRB Cat# JCRB1819, RRID:CVCL_YQ49)

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   Results 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).

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   Results from LimitationRecognizer: We detected the following sentences addressing limitations in the study:

   Reporter assays, including ours, also have limitations. Our reporter utilizes the CoV 3CLpro expressed alone; during a CoV infection the protease is only one of dozens of viral proteins present, and any inhibitors that may affect cross-viral protein interactions would be missed. Additionally, CoV infection induces significant cellular membrane rearrangements that transfection of the protease alone does not. Thus, the subcellular access of therapeutic compounds to the viral protease may fail to be reflected in our assay, and the effects of an identified protease inhibitor could significantly differ when applied to authentic viral infection. Finally, although this plasmid-based expression presents many advantages, it also necessitates further screen hit testing in the context of coronavirus infection. Although the correlation between our reporter and inhibition of viral infection was appreciable with the drug GC376, testing of more inhibitors is required to make generalizable correlations between our reporter assay and viral infection readouts. To have the greatest impact on the COVID-19 pandemic, an effective SARS-CoV-2 antiviral needs to be identified as early as possible. Countries around the world have taken drastic and necessary steps to limit the spread of virus, yet infection rates continue to rise in some. An antiviral treatment is unlikely to stop the spread of infection, but it is likely to limit the mortality associated with SARS-CoV-2 infection. It is our hope that ...

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   Results from TrialIdentifier: No clinical trial numbers were referenced.

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   Results from Barzooka: We found bar graphs of continuous data. We recommend replacing bar graphs with more informative graphics, as many different datasets can lead to the same bar graph. The actual data may suggest different conclusions from the summary statistics. For more information, please see Weissgerber et al (2015).

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   Results from JetFighter: We did not find any issues relating to colormaps.

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   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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   Read the original source
3. 

   Version published to 10.1101/2020.06.24.169565v1 on bioRxiv

   Jun 24, 2020