Viruses within the Poxvirus, Asfarvirus and Iridovirus families carry large double-stranded DNA genomes, replicate in the cytoplasm and infect either invertebrates or vertebrates. A number of these viruses are infectious disease agents for man or domestic and wild animals which is now impressingly demonstrated by the ongoing global monkeypox outbreak. Several poxvirus species are used as vectors for vaccination or therapeutics. The topics to be covered are virus entry, gene expression and proteomics, DNA replication and genomics, virus assembly and release, immunity and immune evasion, virus-host interactions, pathogenesis, vaccines and antivirals, diagnostics, virus vectors, virus oncolytics.