Reverse genetics with a full-length infectious cDNA of severe acute respiratory syndrome coronavirus

Boyd Yount; Kristopher M. Curtis; Elizabeth A. Fritz; Lisa E. Hensley; Peter B. Jahrling; Erik Prentice; Mark R. Denison; Thomas W. Geisbert; Ralph S. Baric

doi:10.1073/pnas.1735582100

Reverse genetics with a full-length infectious cDNA of severe acute respiratory syndrome coronavirus

Boyd Yount, Kristopher M. Curtis, Elizabeth A. Fritz, Lisa E. Hensley, Peter B. Jahrling, Erik Prentice, Mark R. Denison, Thomas W. Geisbert, Ralph S. Baric

Research output: Contribution to journal › Article › peer-review

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Abstract

A previously undescribed coronavirus (CoV) is the etiologic agent responsible for severe acute respiratory syndrome (SARS). Using a panel of contiguous cDNAs that span the entire genome, we have assembled a full-length cDNA of the SARS-CoV Urbani strain, and have rescued molecularly cloned SARS viruses (infectious clone SARS-CoV) that contained the expected marker mutations inserted into the component clones. Recombinant viruses replicated as efficiently as WT virus and both were inhibited by treatment with the cysteine proteinase inhibitor (2S,3S)-transepoxysuccinyl-L-leucylamido-3-methylbutane ethyl ester. In addition, subgenomic transcripts were initiated from the consensus sequence ACGAAC in both the WT and infectious clone SARS-CoV. Availability of a SARS-CoV full-length cDNA provides a template for manipulation of the viral genome, allowing for the rapid and rational development and testing of candidate vaccines and therapeutics against this important human pathogen.

Original language	English (US)
Pages (from-to)	12995-13000
Number of pages	6
Journal	Proceedings of the National Academy of Sciences of the United States of America
Volume	100
Issue number	22
DOIs	https://doi.org/10.1073/pnas.1735582100
State	Published - Oct 28 2003
Externally published	Yes

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Yount, B., Curtis, K. M., Fritz, E. A., Hensley, L. E., Jahrling, P. B., Prentice, E., Denison, M. R., Geisbert, T. W., & Baric, R. S. (2003). Reverse genetics with a full-length infectious cDNA of severe acute respiratory syndrome coronavirus. Proceedings of the National Academy of Sciences of the United States of America, 100(22), 12995-13000. https://doi.org/10.1073/pnas.1735582100

Yount, B, Curtis, KM, Fritz, EA, Hensley, LE, Jahrling, PB, Prentice, E, Denison, MR, Geisbert, TW & Baric, RS 2003, 'Reverse genetics with a full-length infectious cDNA of severe acute respiratory syndrome coronavirus', Proceedings of the National Academy of Sciences of the United States of America, vol. 100, no. 22, pp. 12995-13000. https://doi.org/10.1073/pnas.1735582100

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abstract = "A previously undescribed coronavirus (CoV) is the etiologic agent responsible for severe acute respiratory syndrome (SARS). Using a panel of contiguous cDNAs that span the entire genome, we have assembled a full-length cDNA of the SARS-CoV Urbani strain, and have rescued molecularly cloned SARS viruses (infectious clone SARS-CoV) that contained the expected marker mutations inserted into the component clones. Recombinant viruses replicated as efficiently as WT virus and both were inhibited by treatment with the cysteine proteinase inhibitor (2S,3S)-transepoxysuccinyl-L-leucylamido-3-methylbutane ethyl ester. In addition, subgenomic transcripts were initiated from the consensus sequence ACGAAC in both the WT and infectious clone SARS-CoV. Availability of a SARS-CoV full-length cDNA provides a template for manipulation of the viral genome, allowing for the rapid and rational development and testing of candidate vaccines and therapeutics against this important human pathogen.",

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AU - Curtis, Kristopher M.

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AU - Jahrling, Peter B.

AU - Prentice, Erik

AU - Denison, Mark R.

AU - Geisbert, Thomas W.

AU - Baric, Ralph S.

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AB - A previously undescribed coronavirus (CoV) is the etiologic agent responsible for severe acute respiratory syndrome (SARS). Using a panel of contiguous cDNAs that span the entire genome, we have assembled a full-length cDNA of the SARS-CoV Urbani strain, and have rescued molecularly cloned SARS viruses (infectious clone SARS-CoV) that contained the expected marker mutations inserted into the component clones. Recombinant viruses replicated as efficiently as WT virus and both were inhibited by treatment with the cysteine proteinase inhibitor (2S,3S)-transepoxysuccinyl-L-leucylamido-3-methylbutane ethyl ester. In addition, subgenomic transcripts were initiated from the consensus sequence ACGAAC in both the WT and infectious clone SARS-CoV. Availability of a SARS-CoV full-length cDNA provides a template for manipulation of the viral genome, allowing for the rapid and rational development and testing of candidate vaccines and therapeutics against this important human pathogen.

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Reverse genetics with a full-length infectious cDNA of severe acute respiratory syndrome coronavirus

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