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      Synthetic recombinant bat SARS-like coronavirus is infectious in cultured cells and in mice.

      Proceedings of the National Academy of Sciences of the United States of America

      Amino Acid Sequence, Animals, Cells, Cultured, Cercopithecus aethiops, Chiroptera, virology, Female, Humans, Membrane Glycoproteins, genetics, Mice, Mice, Inbred BALB C, Molecular Sequence Data, Recombinant Proteins, Recombination, Genetic, Respiratory Mucosa, SARS Virus, isolation & purification, physiology, Severe Acute Respiratory Syndrome, Spike Glycoprotein, Coronavirus, Vero Cells, Viral Envelope Proteins, Virus Replication, Zoonoses, transmission

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          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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