Vaccine Development Against COVID-19 Prior to Pandemic Outbreaks, Using in vitro Evolution and Reverse Genetics

Abstract

Developing vaccines against COVID-19 during or after future pandemic outbreaks will be too slow to ‎save lives. The opinion presented here is that we can anticipate the virus genomic structures that will ‎emerge and cause future pandemics; therefore, we can preemptively design a vaccine prior to such a ‎pandemic. This can be done through in vitro evolution, using the open reading frames of the genes ‎from SARS-CoV-2 that code for the immunodominant epitopes of the virus (i.e., N, M, S, and E), as a ‎template for in vitro evolution using DNA shuffling techniques. The resulting recombinant library will ‎be subcloned in lentiviral integrating vectors and be transfected in cell lines that are susceptible for ‎SARS-CoV2, which will be allowed to secrete the corresponding recombinant virus-like particles (VLPs). ‎The resulting VLPs could be tested for protection against mutant viral particles using reverse genetics. ‎Stable, transgenic cell lines could then be generated from the VLP vaccines most protective against the ‎highly pathogenic recombinant viruses. These cell lines could be expanded in large bioreactors to be ‎assembled and shipped to the site(s) of pandemic outbreaks as they happened. This would be ‎expected to prevent deaths during pandemic outbreaks.‎