Broad and Differential Animal Angiotensin-Converting Enzyme 2 Receptor Usage by SARS-CoV-2
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Abstract
SARS-CoV-2 uses human ACE2 as a primary receptor for host cell entry. Viral entry mediated by the interaction of ACE2 with spike protein largely determines host range and is the major constraint to interspecies transmission. We examined the receptor activity of 14 ACE2 orthologs and found that wild-type and mutant SARS-CoV-2 lacking the furin cleavage site in S protein could utilize ACE2 from a broad range of animal species to enter host cells. These results have important implications in the natural hosts, interspecies transmission, animal models, and molecular basis of receptor binding for SARS-CoV-2.
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SciScore for 10.1101/2020.04.19.048710: (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 All growth medium was supplemented with 10% fetal bovine serum (FBS), 110 mg/L sodium pyruvate, and 4.5 g/L D-glucose. β-actin antibody and C9 antibody were purchased from Sigma (A2228) and SANTA CRUZ (sc-57432), respectively. β-actinsuggested: (Sigma-Aldrich Cat# A2228, RRID:AB_476697)C9suggested: NoneA polyclonal antibody against human ACE2 and anti-IDE polyclonal antibody were purchased from R&D Systems (catalog No. AF933 and AF2496, respectively). human …SciScore for 10.1101/2020.04.19.048710: (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 All growth medium was supplemented with 10% fetal bovine serum (FBS), 110 mg/L sodium pyruvate, and 4.5 g/L D-glucose. β-actin antibody and C9 antibody were purchased from Sigma (A2228) and SANTA CRUZ (sc-57432), respectively. β-actinsuggested: (Sigma-Aldrich Cat# A2228, RRID:AB_476697)C9suggested: NoneA polyclonal antibody against human ACE2 and anti-IDE polyclonal antibody were purchased from R&D Systems (catalog No. AF933 and AF2496, respectively). human ACE2suggested: Noneanti-IDEsuggested: NoneExperimental Models: Cell Lines Sentences Resources Cell lines and antibodies: 293T cells and Lenti-X 293T cells were cultured in Dulbecco’s modified Eagle’s medium (DMEM; Gibco) (57). 293Tsuggested: NoneBriefly, HEK293T cells were transfected with plasmid encoding ACE2 with Lipofectamine 2000 (Invitrogen). HEK293Tsuggested: NoneSoftware and Algorithms Sentences Resources Construction of plasmids expressing S, S1 and RBD of SARS-CoV-2: The nucleotide sequence of SARS-CoV-2 S gene was retrieved from NCBI database (isolate Wuhan-Hu-1, GenBank No. MN908947). NCBIsuggested: (NCBI, RRID:SCR_006472)Phylogenetic tree was constructed based on the nucleotide sequences of animal ACE2 using the neighbor-joining algorithm implemented in MEGA X. MEGAsuggested: (Mega BLAST, RRID:SCR_011920)The models were then aligned and compared to the intact SARS-CoV-2 RBD/ ACE2 complex in PyMOL (The PyMOL Molecular Graphics System, Version 2.0 Schrödinger, LLC). PyMOLsuggested: (PyMOL, RRID:SCR_000305)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: 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.
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