Generation of SARS-CoV-2 Spike Variants

rVSVs expressing the SARS-CoV S (GenBank: AY278554.2) (termed as rVSV-SARS), the SARS-CoV-2 prototype strain S (GenBank: MN908947) (termed as rVSV-SARS2), SARS-CoV-2 VOCs S, or 55 SARS-CoV-2 S with different single point mutations were generated as previously described (33 (link)). Briefly, SARS-CoV or SARS-CoV-2 genes encoding for the S protein were cloned separately into the pCAG eukaryotic expression plasmid (Addgene). These genes had an 18–amino acid C-terminal truncation. rSARS-CoV and rSARS-CoV2 were rescued by VSVdG-EGFP-G (Addgene, 31842) from the Vero E6 cells transfected with plasmids pCAG-SARS1-Sdel18 and pCAG-SARS2-Sdel18 (48 (link)), respectively. Supernatants were harvested and purified by Capto Core 700 (Cytiva) multimodal chromatography. The viral particles were collected in the column flowthrough.

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Xiong H., Sun H., Wang S., Yuan L., Liu L., Zhu Y., Zhang J., Huang Y., Qi R., Jiang Y., Ma J., Zhou M., Ma Y., Fu R., Yan S., Yue M., Wu Y., Wei M., Wang Y., Li T., Wang Y., Zheng Z., Yu H., Cheng T., Li S., Yuan Q., Zhang J., Guan Y., Zheng Q., Zhang T, & Xia N. (2022). The neutralizing breadth of antibodies targeting diverse conserved epitopes between SARS-CoV and SARS-CoV-2. Proceedings of the National Academy of Sciences of the United States of America, 119(34), e2204256119.

Publication 2022

VSVdG-EGFP-G Capto Core 700

Amino acid Chromatography Eukaryotic Genes Multimodal Plasmids Point mutations S protein Sars Sars cov Sars2 Strain Vero cells Viral particles

Corresponding Organization :

Other organizations : Xiamen University, Shantou University, University of Hong Kong, Chinese Academy of Medical Sciences & Peking Union Medical College

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Variable analysis

independent variables

RVSVs expressing the SARS-CoV S (GenBank: AY278554.2) (termed as rVSV-SARS)
The SARS-CoV-2 prototype strain S (GenBank: MN908947) (termed as rVSV-SARS2)
SARS-CoV-2 VOCs S
55 SARS-CoV-2 S with different single point mutations

dependent variables

Not explicitly mentioned

control variables

18–amino acid C-terminal truncation of SARS-CoV or SARS-CoV-2 genes encoding for the S protein
Vero E6 cells transfected with plasmids pCAG-SARS1-Sdel18 and pCAG-SARS2-Sdel18 for rescuing rSARS-CoV and rSARS-CoV2 respectively
Supernatants harvested and purified by Capto Core 700 (Cytiva) multimodal chromatography

controls

Positive control: not mentioned
Negative control: not mentioned

Annotations

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