Mutagenesis of the of SARS-Coronavirus-2 Spike Gene

Coronavirus disease 2019 (COVID-19), caused by the SARS-CoV-2 virus, has become a global pandemic resulting in over 6 million deaths worldwide. The virus binds to host cells through its spike (S) attachment protein, composed of two subunits: S1 and S2. The S protein is a target for vaccine development, with neutralizing antibodies blocking ACE2 binding to prevent infection. However, SARS-CoV-2 continues to evolve and accumulate mutations in the S protein, including L452R, T478K, D614G, P681R, and D950N, which can increase infectivity and evade host immune functions. To study the functions of these mutations, we randomly combined them in the alpha variant S gene using primers of differing concentrations, generating a library of transformants with unique combinations of mutations. Sequencing to confirm mutations is pending, and the future goal of this project is to use safe-to-use virus-like particles psuedotyped with the mutated SARS-CoV-2 S proteins which will be used in entry and immune cell deregulation assays