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Synthetic recombinant bat SARS-like coronavirus is infectious in cultured cells and in mice
Michelle M. Becker, Rachel L. Graham, Eric F. Donaldson, +7, and Mark R. Denison rbaric@email.unc.eduAuthors Info & Affiliations
Edited by Peter Palese, Mount Sinai School of Medicine, New York, NY, and approved October 14, 2008
December 16, 2008
105 (50) 19944-19949
https://doi.org/10.1073/pnas.0808116105
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Vol. 105 | No. 50
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Abstract
Defining prospective pathways by which zoonoses evolve and emerge as human pathogens is critical for anticipating and controlling both natural and deliberate pandemics. However, predicting tenable pathways of animal-to-human movement has been hindered by challenges in identifying reservoir species, cultivating zoonotic organisms in culture, and isolating full-length genomes for cloning and genetic studies. The ability to design and recover pathogens reconstituted from synthesized cDNAs has the potential to overcome these obstacles by allowing studies of replication and pathogenesis without identification of reservoir species or cultivation of primary isolates. Here, we report the design, synthesis, and recovery of the largest synthetic replicating life form, a 29.7-kb bat severe acute respiratory syndrome (SARS)-like coronavirus (Bat-SCoV), a likely progenitor to the SARS-CoV epidemic. To test a possible route of emergence from the noncultivable Bat-SCoV to human SARS-CoV, we designed a consensus Bat-SCoV genome and replaced the Bat-SCoV Spike receptor-binding domain (RBD) with the SARS-CoV RBD (Bat-SRBD). Bat-SRBD was infectious in cell culture and in mice and was efficiently neutralized by antibodies specific for both bat and human CoV Spike proteins. Rational design, synthesis, and recovery of hypothetical recombinant viruses can be used to investigate mechanisms of transspecies movement of zoonoses and has great potential to aid in rapid public health responses to known or predicted emerging microbial threats.
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Data Availability
Data deposition: The sequences reported in this paper have been deposited in the GenBank database (accession nos. FJ211859 and FJ211860).
Acknowledgments.
We thank XiaoTao Lu and Sunny Lee for technical assistance, Susan Burkett for maintenance of the HAE cultures, Perry Myrick for immunofluorescence assays, and the University of North Carolina Cystic Fibrosis Tissue Culture Core for HAE cells. M.M.B., R.L.G., R.S.B., and M.R.D. are supported by the National Institute of Allergy and Infectious Diseases Public Health Service Award P01 AI59943. Additional support was provided by Public Health Service Award CA68485 to the Vanderbilt University DNA Sequencing Shared Resource of the Vanderbilt–Ingram Cancer Center. The Baric laboratory is supported by the Gillings Innovation Fund.
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