Recombinant Lloviu virus (rLLOV-ZsGreen)

Many potentially zoonotic viruses that are closely related to pathogens causing recent human epidemics and pandemics remain understudied because they have not yet been cultured. While virus isolation from wildlife poses challenges, reverse genetic systems can facilitate the rescue of these viruses, although this process is hindered by frequently missing genomic ends. Here we utilize BSL2 minigenome systems to test the ability of heterologous filovirus sequences to complement the missing genomic ends of Lloviu virus (LLOV), a previously uncultured filovirus endemic in Europe. Leveraging these data, we rescued a recombinant infectious LLOV that allowed us to investigate cellular tropism in human primary cells and potential antiviral therapies. Rescue of uncultured zoonotic viruses of pathogenic concern in this manner represents a valuable tool in our arsenal in pandemic preparedness.