The SARS-CoV-2 Transcriptome and the Dynamics of the S Gene Furin Cleavage Site in Primary Human Airway Epithelia
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Abstract
Polarized human airway epithelia at an air-liquid interface (HAE-ALI) are an in vitro model that supports efficient infection of SARS-CoV-2. The spike (S) protein of SARS-CoV-2 contains a furin cleavage site (FCS) at the boundary of the S1 and S2 domains which distinguishes it from SARS-CoV.
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SciScore for 10.1101/2021.02.03.429670: (What is this?)
Please note, not all rigor criteria are appropriate for all manuscripts.
Table 1: Rigor
Institutional Review Board Statement IACUC: A biosafety protocol to work on SARS-CoV-2 infection in the biosafety level-3 (BSL3) lab was approved by the Institutional Biosafety Committee of the University of Kansas Medical Center. 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 The virus used for infections of HAE-ALI was propagated once in Vero-E6 cells, designated as P1 passage. Vero-E6suggested: NoneHAE-ALI cultures: Primary HAE-ALI cultures, lots of B2-20, B3-20, B4-20, B9-20, B15-20, and B16-20, were prepared from bronchial … SciScore for 10.1101/2021.02.03.429670: (What is this?)
Please note, not all rigor criteria are appropriate for all manuscripts.
Table 1: Rigor
Institutional Review Board Statement IACUC: A biosafety protocol to work on SARS-CoV-2 infection in the biosafety level-3 (BSL3) lab was approved by the Institutional Biosafety Committee of the University of Kansas Medical Center. 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 The virus used for infections of HAE-ALI was propagated once in Vero-E6 cells, designated as P1 passage. Vero-E6suggested: NoneHAE-ALI cultures: Primary HAE-ALI cultures, lots of B2-20, B3-20, B4-20, B9-20, B15-20, and B16-20, were prepared from bronchial airway epithelial cells isolated from various donors. B16-20suggested: NoneSoftware and Algorithms Sentences Resources The purified DNA samples were quantified on a microplate reader (Synergy LX, BioTek, Winooski, VT), and 500 ng of each DNA sample (20 ng/µl) was sent for PCR amplicon-seq (AMPLICON-EZ) at GENEWIZ, Inc. (South Plainfield, NJ) GENEWIZsuggested: (GENEWIZ, RRID:SCR_003177)Bioinformatic analyses: Bioinformaticsuggested: (QFAB Bioinformatics, RRID:SCR_012513)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: We detected the following sentences addressing limitations in the study:Since the FCS is a key motif related to virulence, it is important to investigate the natural occurrence rate of the FCS region deletions, possibility or limitation of their human-to-human transmission, as well as their pathogenicity. Several studies tried to screen the FCS region deletions from patients-derived SARS-CoV-2. As discussed above, screening of 27 SARS-CoV-2 positive clinical specimens, including one specimen that had FCS deletions identified after passaging in Vero-E6 cells, failed to detect any FCS deletions (33). However, one study detected the 675QTQTN679-deleted mutants (mut-del2) in 3 of 68 SARS-CoV-2 positive clinical specimens (32). In another detection of 51 SARS-CoV-2 positive patient specimens, although a high rate of 52.9% and 82.4% of the positive clinical samples contained the FCS upstream motif (661ECDIPIGAG669) and the PRRA deletions, respectively, the mutant population is at a very low level (0.33% ±1.17% for FCS upstream motif deletion and 1.12% ±1.21% for PRRA deletion) (51), arguing the infectivity and transmissibility of these mutants. Along with the usages of antibody drugs and the wide inoculation of the vaccine, which target the S protein, the virus may undergo further mutations under the pressure of human immune response. Supervision and screening the mutations in the S protein gene in clinical specimens is extremely important to identify the escaped isolates which may increase or decrease infectivity and transmissibility. Apparently, the ...
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.
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- No protocol registration statement was detected.
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