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Multi-color super-resolution imaging to study human coronavirus RNA during cellular infection

View ORCID ProfileJiarui Wang, Mengting Han, Anish R. Roy, Haifeng Wang, Leonhard Möckl, Leiping Zeng, View ORCID ProfileW. E. Moerner, View ORCID ProfileLei S. Qi
doi: https://doi.org/10.1101/2021.06.09.447760
Jiarui Wang
1Department of Chemistry, Chemical and Systems Biology, and ChEM-H Stanford University, Stanford, California, 94305 U.S.A.
2Department of Developmental Biology, Chemical and Systems Biology, and ChEM-H Stanford University, Stanford, California, 94305 U.S.A.
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  • ORCID record for Jiarui Wang
Mengting Han
3Departments of Bioengineering, Chemical and Systems Biology, and ChEM-H Stanford University, Stanford, California, 94305 U.S.A.
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Anish R. Roy
1Department of Chemistry, Chemical and Systems Biology, and ChEM-H Stanford University, Stanford, California, 94305 U.S.A.
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Haifeng Wang
3Departments of Bioengineering, Chemical and Systems Biology, and ChEM-H Stanford University, Stanford, California, 94305 U.S.A.
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Leonhard Möckl
1Department of Chemistry, Chemical and Systems Biology, and ChEM-H Stanford University, Stanford, California, 94305 U.S.A.
4Max Planck Institute for the Science of Light, Erlangen, Germany
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Leiping Zeng
3Departments of Bioengineering, Chemical and Systems Biology, and ChEM-H Stanford University, Stanford, California, 94305 U.S.A.
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W. E. Moerner
1Department of Chemistry, Chemical and Systems Biology, and ChEM-H Stanford University, Stanford, California, 94305 U.S.A.
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  • For correspondence: slqi@stanford.edu wmoerner@stanford.edu
Lei S. Qi
3Departments of Bioengineering, Chemical and Systems Biology, and ChEM-H Stanford University, Stanford, California, 94305 U.S.A.
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  • For correspondence: slqi@stanford.edu wmoerner@stanford.edu
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SUMMARY

The severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) is the third human coronavirus within 20 years that gave rise to a life-threatening disease and the first to reach pandemic spread. To make therapeutic headway against current and future coronaviruses, the biology of coronavirus RNA during infection must be precisely understood. Here, we present a robust and generalizable framework combining high-throughput confocal and super-resolution microscopy imaging to study coronavirus infection at the nanoscale. Employing the model human coronavirus HCoV-229E, we specifically labeled coronavirus genomic RNA (gRNA) and double-stranded RNA (dsRNA) via multicolor RNA-immunoFISH and visualized their localization patterns within the cell. The exquisite resolution of our approach uncovers a striking spatial organization of gRNA and dsRNA into three distinct structures and enables quantitative characterization of the status of the infection after antiviral drug treatment. Our approach provides a comprehensive framework that supports investigations of coronavirus fundamental biology and therapeutic effects.

Competing Interest Statement

The authors have declared no competing interest.

Footnotes

  • Revised version of the manuscript with new data to co-image Spike protein and viral genome RNA (Figure 3)

Copyright 
The copyright holder for this preprint is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. It is made available under a CC-BY-NC-ND 4.0 International license.
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Posted January 13, 2022.
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Multi-color super-resolution imaging to study human coronavirus RNA during cellular infection
Jiarui Wang, Mengting Han, Anish R. Roy, Haifeng Wang, Leonhard Möckl, Leiping Zeng, W. E. Moerner, Lei S. Qi
bioRxiv 2021.06.09.447760; doi: https://doi.org/10.1101/2021.06.09.447760
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Multi-color super-resolution imaging to study human coronavirus RNA during cellular infection
Jiarui Wang, Mengting Han, Anish R. Roy, Haifeng Wang, Leonhard Möckl, Leiping Zeng, W. E. Moerner, Lei S. Qi
bioRxiv 2021.06.09.447760; doi: https://doi.org/10.1101/2021.06.09.447760

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