Directed Evolution of Adenoviruses.

The ability to evolve viruses in cell culture in the face of selective pressure is an invaluable method to elucidate the molecular mechanisms of synthetic or natural antivirals, expand tropism, or alter virulence. Recently, mutations to the human adenovirus polymerase that reduce replicative fidelity were described, and we have incorporated one of these mutations into the pol gene of a conditionally replicating human adenovirus serotype 5 (HAdV-5)-based vector (Uil et al., Nucleic Acids Res 39:e30, 2011; Myers et al., J Virol 87:6047-6050, 2013). Here, we describe methods to evolve this virus (HAdV-5.polF421Y) under selective pressure from antivirals to identify their mechanisms of action.