Host-Virus Chimeric Events in SARS-CoV-2-Infected Cells Are Infrequent and Artifactual

  1. Bingyu Yan
  2. Srishti Chakravorty
  3. Carmen Mirabelli
  4. Luopin Wang
  5. Jorge L. Trujillo-Ochoa
  6. Daniel Chauss
  7. Dhaneshwar Kumar
  8. Michail S. Lionakis
  9. Matthew R. Olson
  10. Christiane E. Wobus
  11. Behdad Afzali
  12. Majid Kazemian

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Abstract

The pathogenic mechanisms underlying SARS-CoV-2, the virus responsible for COVID-19, are not fully understood. In particular, relatively little is known about the reasons some individuals develop life-threatening or persistent COVID-19.

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  1. Version published to 10.1128/jvi.00294-21
  2. ScreenIT

    SciScore for 10.1101/2021.02.17.431704: (What is this?)

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

    Table 1: Rigor

    Institutional Review Board StatementIACUC: Experiments using SARS-CoV-2 were performed at the University of Michigan under Biosafety Level 3 (BSL3) protocols in compliance with containment procedures in laboratories approved for use by the University of Michigan Institutional Biosafety Committee (IBC) and Environment, Health & Safety (EHS).
    Randomizationnot detected.
    Blindingnot detected.
    Power Analysisnot detected.
    Sex as a biological variablenot detected.
    Cell Line Authenticationnot detected.

    Table 2: Resources

    Experimental Models: Cell Lines
    SentencesResources
    SARS coronavirus 2 (SARS-CoV-2), isolate USA-WA1/2020 (NR-52281) was obtained from Bei resources and was propagated in Vero E6 cells in DMEM supplemented with 2% FBS, 4.5 g/L D-glucose, 4 mM L-glutamine, 10 mM Non-Essential Amino Acids, 1 mM Sodium Pyruvate and 10 mM HEPES.
    Vero E6
    suggested: RRID:CVCL_XD71)
    Fifty thousand Calu-3 were seeded in 48 well plate and allowed to form 80% confluent monolayer.
    Calu-3
    suggested: None
    Software and Algorithms
    SentencesResources
    Host-virus chimeric read analysis: The raw sequencing files were download from the Sequence Read Archive (SRA) as shown in the Table S1.
    Sequence Read Archive
    suggested: (DDBJ Sequence Read Archive, RRID:SCR_001370)
    Sequencing reads were aligned as single end to the chimeric genome of human (hg38) and SARS-CoV2 (NC_045512.2) using STAR aligner (v2.7.7a).
    STAR
    suggested: (STAR, RRID:SCR_015899)
    To examine the genomic features of the HVC reads, HOMER (v4.11) annotatePeaks.pl was used to annotate the HVC junctions and the corresponding RNA-seq library.
    HOMER
    suggested: (HOMER, RRID:SCR_010881)
    In brief, reads in each RNA-seq library were converted to genomic regions by bamTobed (bedtools, v2.30.0) and the unique regions were kept using the following command”sort -k1,1 -k2,2n | uniq”.
    bedtools
    suggested: (BEDTools, RRID:SCR_006646)

    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: 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.

    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 10.1101/2021.02.17.431704 on bioRxiv