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Significance All known membrane-enveloped RNA viruses have capsid proteins that encase their RNA genomes. This paper shows that infectious, membrane-enveloped, virus-like vesicles with RNA genomes can evolve in vitro to grow to high titers without a capsid protein. The infectious vesicles are apparently generated from RNA replication factories called spherules that bud from the cell surface. They evolve in vitro to bud with high efficiency through the acquisition of multiple mutations in the non-structural replicase proteins. One mutation generates a critical motif found in many viral structural proteins. This motif is involved in recruiting cellular machinery to drive efficient budding. Prior to the evolution of capsid proteins, primitive RNA viruses may have used this budding mechanism.

In vitro evolution of high-titer, virus-like vesicles containing a single structural protein