Large-scale packaging of alphavirus replicons is accomplished by co-transfecting
host cells with three RNA molecules: (1) an alphavirus replicon comprising a sequence
encoding a heterologous protein; (2) a helper RNA encoding a capsid protein and
(3) a helper RNA encoding two alphavirus glycoproteins. The helper RNAs contain
cis-acting elements that allow efficient replication of the helper RNAs and their
packaging into viral particles as well as packaging of replicon genomes. These
populations of viral particles can be propagated at high titers in cell culture
by infecting cells at high multiplicity. Propagation of packaged replicons at an
escalating scale is useful for large-scale production of recombinant proteins and/or vaccine.