SARS-CoV-2 and Three Related Coronaviruses Utilize Multiple ACE2 Orthologs and Are Potently Blocked by an Improved ACE2-Ig
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Abstract
The severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) is the etiological agent of the currently uncontrolled coronavirus disease 2019 (COVID-19) pandemic. It is important to study the host range of SARS-CoV-2, because some domestic species might harbor the virus and transmit it back to humans. In addition, insight into the ability of SARS-CoV-2 and SARS-like viruses to utilize animal orthologs of the SARS-CoV-2 receptor ACE2 might provide structural insight into improving ACE2-based viral entry inhibitors. In this study, we found that ACE2 orthologs of a wide range of domestic and wild animals can support cell entry of SARS-CoV-2 and three related coronaviruses, providing insights into identifying animal hosts of these viruses. We also developed recombinant ACE2-Ig proteins that are able to potently block these viral infections, providing a promising approach to developing antiviral proteins broadly effective against these distinct coronaviruses.
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SciScore for 10.1101/2020.04.10.032342: (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 Cells were then stained with 5 μg/mL RBD-Fc proteins at 37 °C for 10 min, washed three times, and then stained with 2 μg/mL Alexa488 conjugated goat anti-mouse IgG secondary antibody (Invitrogen, Cat. No. A-11001) at room temperature for 20 min. anti-mouse IgGsuggested: (Thermo Fisher Scientific Cat# A-11001, RRID:AB_2534069)ACE2-S-tag expression was detected by using 6.2, a mouse anti-S-tag monoclonoal antibody (Invitrogen, Cat. No. MA1-981), and an … SciScore for 10.1101/2020.04.10.032342: (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 Cells were then stained with 5 μg/mL RBD-Fc proteins at 37 °C for 10 min, washed three times, and then stained with 2 μg/mL Alexa488 conjugated goat anti-mouse IgG secondary antibody (Invitrogen, Cat. No. A-11001) at room temperature for 20 min. anti-mouse IgGsuggested: (Thermo Fisher Scientific Cat# A-11001, RRID:AB_2534069)ACE2-S-tag expression was detected by using 6.2, a mouse anti-S-tag monoclonoal antibody (Invitrogen, Cat. No. MA1-981), and an HRP-conjugated goat anti-mouse IgG Fc secondary antibody (Invitrogen, Cat. No. 31437). anti-S-tagsuggested: (Thermo Fisher Scientific Cat# MA1-981, RRID:AB_347008)Spike-C9-tag expression was then detected by using 1D4, a mouse anti-C9-tag monoclonal antibody (Invitrogen, Cat. No. MA1-722), and the HRP-conjugated goat anti-mouse IgG Fc secondary antibody (Invitrogen, Cat. No. 31437). anti-C9-tagsuggested: NoneExperimental Models: Cell Lines Sentences Resources 293T-based stable cells expressing human ACE2 were maintained under the same culture condition as 293T, except that 3 μg/mL of puromycin was added to the growth medium. 293T-basedsuggested: None293F cells for recombinant protein production was generously provided by Dr. Yu J. 293Fsuggested: RRID:CVCL_D615)Production of reporter retroviruses pseudotyped with coronavirus spike proteins or VSV-G: 293T cells were seeded at 30% density in 150 mm dish at 12-15 hours before transfection. 293Tsuggested: NoneSoftware and Algorithms Sentences Resources Statistical analyses were performed using two-sided two-sample Student’s t-test using GraphPad Prism 6.0 software when applicable. GraphPad Prismsuggested: (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 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).
Results from JetFighter: Please consider improving the rainbow (“jet”) colormap(s) used on page 12. 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:- 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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