Generation of Recombinant SARS‐CoV‐2 Using a Bacterial Artificial Chromosome

SARS‐CoV‐2, the causative agent of COVID‐19, has been responsible for a million deaths worldwide as of September 2020. At the time of this writing, there are no available US FDA−approved therapeutics for the treatment of SARS‐CoV‐2 infection. Here, we describe a detailed protocol to generate recombinant (r)SARS‐CoV‐2 using reverse‐genetics approaches based on the use of a bacterial artificial chromosome (BAC). This method will allow the production of mutant rSARS‐CoV‐2—which is necessary for understanding the function of viral proteins, viral pathogenesis and/or transmission, and interactions at the virus‐host interface—and attenuated SARS‐CoV‐2 to facilitate the discovery of effective countermeasures to control the ongoing SARS‐CoV‐2 pandemic. © 2020 Wiley Periodicals LLC. Basic Protocol : Generation of recombinant SARS‐CoV‐2 using a bacterial artificial chromosome Support Protocol : Validation and characterization of rSARS‐CoV‐2