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SARS CoV-2 Nucleoprotein Enhances the Infectivity of Lentiviral Spike Particles

The establishment of SARS CoV-2 spike-pseudotyped lentiviral (LV) systems has enabled the rapid identification of entry inhibitors and neutralizing agents, alongside allowing for the study of this emerging pathogen in BSL-2 level facilities. While such frameworks recapitulate the cellular entry proc...

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Autores principales: Mishra, Tarun, Sreepadmanabh, M., Ramdas, Pavitra, Sahu, Amit Kumar, Kumar, Atul, Chande, Ajit
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Frontiers Media S.A. 2021
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8102828/
https://www.ncbi.nlm.nih.gov/pubmed/33968806
http://dx.doi.org/10.3389/fcimb.2021.663688
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author Mishra, Tarun
Sreepadmanabh, M.
Ramdas, Pavitra
Sahu, Amit Kumar
Kumar, Atul
Chande, Ajit
author_facet Mishra, Tarun
Sreepadmanabh, M.
Ramdas, Pavitra
Sahu, Amit Kumar
Kumar, Atul
Chande, Ajit
author_sort Mishra, Tarun
collection PubMed
description The establishment of SARS CoV-2 spike-pseudotyped lentiviral (LV) systems has enabled the rapid identification of entry inhibitors and neutralizing agents, alongside allowing for the study of this emerging pathogen in BSL-2 level facilities. While such frameworks recapitulate the cellular entry process in ACE2+ cells, they are largely unable to factor in supplemental contributions by other SARS CoV-2 genes. To address this, we performed an unbiased ORF screen and identified the nucleoprotein (N) as a potent enhancer of spike-pseudotyped LV particle infectivity. We further demonstrate that the spike protein is better enriched in virions when the particles are produced in the presence of N protein. This enrichment of spike renders LV particles more infectious as well as less vulnerable to the neutralizing effects of a human IgG-Fc fused ACE2 microbody. Importantly, this improvement in infectivity is observed with both wild-type spike protein as well as the D614G mutant. Our results hold important implications for the design and interpretation of similar LV pseudotyping-based studies.
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spelling pubmed-81028282021-05-08 SARS CoV-2 Nucleoprotein Enhances the Infectivity of Lentiviral Spike Particles Mishra, Tarun Sreepadmanabh, M. Ramdas, Pavitra Sahu, Amit Kumar Kumar, Atul Chande, Ajit Front Cell Infect Microbiol Cellular and Infection Microbiology The establishment of SARS CoV-2 spike-pseudotyped lentiviral (LV) systems has enabled the rapid identification of entry inhibitors and neutralizing agents, alongside allowing for the study of this emerging pathogen in BSL-2 level facilities. While such frameworks recapitulate the cellular entry process in ACE2+ cells, they are largely unable to factor in supplemental contributions by other SARS CoV-2 genes. To address this, we performed an unbiased ORF screen and identified the nucleoprotein (N) as a potent enhancer of spike-pseudotyped LV particle infectivity. We further demonstrate that the spike protein is better enriched in virions when the particles are produced in the presence of N protein. This enrichment of spike renders LV particles more infectious as well as less vulnerable to the neutralizing effects of a human IgG-Fc fused ACE2 microbody. Importantly, this improvement in infectivity is observed with both wild-type spike protein as well as the D614G mutant. Our results hold important implications for the design and interpretation of similar LV pseudotyping-based studies. Frontiers Media S.A. 2021-04-23 /pmc/articles/PMC8102828/ /pubmed/33968806 http://dx.doi.org/10.3389/fcimb.2021.663688 Text en Copyright © 2021 Mishra, Sreepadmanabh, Ramdas, Sahu, Kumar and Chande https://creativecommons.org/licenses/by/4.0/This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.
spellingShingle Cellular and Infection Microbiology
Mishra, Tarun
Sreepadmanabh, M.
Ramdas, Pavitra
Sahu, Amit Kumar
Kumar, Atul
Chande, Ajit
SARS CoV-2 Nucleoprotein Enhances the Infectivity of Lentiviral Spike Particles
title SARS CoV-2 Nucleoprotein Enhances the Infectivity of Lentiviral Spike Particles
title_full SARS CoV-2 Nucleoprotein Enhances the Infectivity of Lentiviral Spike Particles
title_fullStr SARS CoV-2 Nucleoprotein Enhances the Infectivity of Lentiviral Spike Particles
title_full_unstemmed SARS CoV-2 Nucleoprotein Enhances the Infectivity of Lentiviral Spike Particles
title_short SARS CoV-2 Nucleoprotein Enhances the Infectivity of Lentiviral Spike Particles
title_sort sars cov-2 nucleoprotein enhances the infectivity of lentiviral spike particles
topic Cellular and Infection Microbiology
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8102828/
https://www.ncbi.nlm.nih.gov/pubmed/33968806
http://dx.doi.org/10.3389/fcimb.2021.663688
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