Contribution of SARS-CoV-2 Accessory Proteins to Viral Pathogenicity in K18 Human ACE2 Transgenic Mice

  1. Jesus A. Silvas
  2. Desarey Morales Vasquez
  3. Jun-Gyu Park
  4. Kevin Chiem
  5. Anna Allué-Guardia
  6. Andreu Garcia-Vilanova
  7. Roy Neal Platt
  8. Lisa Miorin
  9. Thomas Kehrer
  10. Anastasija Cupic
  11. Ana S. Gonzalez-Reiche
  12. Harm van Bakel
  13. Adolfo García-Sastre
  14. Tim Anderson
  15. Jordi B. Torrelles
  16. Chengjin Ye
  17. Luis Martinez-Sobrido

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Abstract

Despite great efforts put forward worldwide to combat the current coronavirus disease 2019 (COVID-19) pandemic, severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) continues to be a human health and socioeconomic threat. Insights into the pathogenesis of SARS-CoV-2 and the contribution of viral proteins to disease outcome remain elusive.

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  1. Version published to 10.1128/jvi.00402-21
  2. ScreenIT

    SciScore for 10.1101/2021.03.09.434696: (What is this?)

    Please note, not all rigor criteria are appropriate for all manuscripts.

    Table 1: Rigor

    Institutional Review Board StatementIACUC: Biosafety (IBC) and Animal Care and Use (IACUC) committees.
    Randomizationnot detected.
    Blindingnot detected.
    Power Analysisnot detected.
    Sex as a biological variableMice: Specific-pathogen-free, 4–8 week-old, female B6.Cg-Tg(K18-ACE2)2Prlmn/J (Stock No: 034860
    Cell Line Authenticationnot detected.

    Table 2: Resources

    Antibodies
    SentencesResources
    Cells were incubated overnight with 1 μg/ml of a SARS-CoV N protein cross-reactive polyclonal antibody at 4°C and a SARS-CoV-2 spike (S) cross-reactive monoclonal antibody (3B4), washed with PBS, and stained with a FITC-labeled goat anti-mouse IgG (1:200) and Rhodamine-labeled goat anti-rabbit IgG (1:200).
    anti-mouse IgG
    suggested: None
    anti-rabbit IgG
    suggested: None
    Experimental Models: Cell Lines
    SentencesResources
    Cells: African green monkey kidney epithelial cells (Vero E6, CRL-1586) were obtained from the American Type Culture Collection (ATCC, Bethesda, MD) and maintained in Dulbecco’s modified Eagle medium (DMEM) supplemented with 5% (v/v)
    Vero E6
    suggested: None
    Experimental Models: Organisms/Strains
    SentencesResources
    Mice: Specific-pathogen-free, 4–8 week-old, female B6.Cg-Tg(K18-ACE2)2Prlmn/J (Stock No: 034860
    B6.Cg-Tg(K18-ACE2)2Prlmn/J
    suggested: RRID:IMSR_JAX:034860)
    K18 hACE2 transgenic mice were purchased from The Jackson Laboratory (Bar Harbor, ME).
    K18 hACE2
    suggested: RRID:IMSR_GPT:T037657)
    Software and Algorithms
    SentencesResources
    Statistical analysis: Statistical analysis was performed using GraphPad Prism 8.3.
    GraphPad Prism
    suggested: (GraphPad Prism, RRID:SCR_002798)

    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).

    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: Please consider improving the rainbow (“jet”) colormap(s) used on page 34. At least one figure is not accessible to readers with colorblindness and/or is not true to the data, i.e. not perceptually uniform.

    Results from rtransparent:
    • No conflict of interest statement was detected. If there are no conflicts, we encourage authors to explicit state so.
    • 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.

    About SciScore

    SciScore is an automated tool that is designed to assist expert reviewers by finding and presenting formulaic information scattered throughout a paper in a standard, easy to digest format. SciScore checks for the presence and correctness of RRIDs (research resource identifiers), and for rigor criteria such as sex and investigator blinding. For details on the theoretical underpinning of rigor criteria and the tools shown here, including references cited, please follow this link.

  3. Version published to 10.1101/2021.03.09.434696v1 on bioRxiv