Demonstration of a TAR-based coronavirus reverse genetics system for the generation of recombinant viruses
Wilson, Natalie R
Harris, Debreiona Y
Smith, Everett Clinton
Scholarship Sewanee 2022 , coronavirus , reverse genetics , transformation-associated recombination , mouse hepatitis virus , viral reverse genetics
Coronaviruses (CoVs), like all viruses, are obligate intracellular parasites, meaning they rely on host molecular machinery (e.g., ribosomes), resources (e.g., nucleotides, amino acids, NTPs), and cellular environment in order to produce progeny. Viral reverse genetics is a key experimental tool for the generation of recombinant viruses that enable the study of viral replication, evolution, and the evaluation of potential therapeutic targets. Herein, we present the first plasmid-launched, transformation-associated recombination (TAR) reverse genetics system for the BSL-2 betacoronavirus, mouse hepatitis virus strain A59 (MHV-A59) and provide proof-of-principle for the utility of this system to generate a panel of recombinant viruses in only a few weeks.