title: Replication-deficient vaccinia virus encoding bacteriophage T7 RNA polymerase for transient gene expression in mammalian cells.
abstract: The vaccinia virus/bacteriophage T7 hybrid transient expression system employs a recombinant vaccinia virus that encodes the T7 RNA polymerase gene, a plasmid vector with a gene of interest regulated by a T7 promoter, and any cell line suitable for infection and transfection. Although high expression in a majority of cells is achieved, the severe cytopathic effects of vaccinia virus and the safety precautions required for use of infectious agents are undesirable features of the system. Here, we report the construction of a highly attenuated and avian host-restricted vaccinia virus recombinant that encodes the T7 RNA polymerase gene (MVA/T7 pol) and demonstrate the use of the virus for transient expression in mammalian cells. MVA/T7 pol has reduced cytopathic effects compared to the previously used replication-competent vaccinia virus, while providing a high level of gene expression in multiple mammalian cell lines.
authors.affiliation: Laboratory of Viral Diseases, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Bethesda, Maryland 20892-0455, USA.
authors.forename: L S
authors.initials: LS
authors.lastname: Wyatt
authors.forename: B
authors.initials: B
authors.lastname: Moss
authors.forename: S
authors.initials: S
authors.lastname: Rozenblatt
mesh_terms: D000818:Animals; D017123:Bacteriophage T7 / Q000201:enzymology*; D002460:Cell Line; D012321:DNA-Directed RNA Polymerases / Q000096:biosynthesis*; D015870:Gene Expression; D007700:Kinetics; D008322:Mammals; D011994:Recombinant Proteins / Q000096:biosynthesis; D011995:Recombination, Genetic; D012449:Safety; D014162:Transfection / Q000379:methods*; D014616:Vaccinia virus / Q000235:genetics*; D014764:Viral Proteins; D014773:Virology / Q000592:standards; D001616:beta-Galactosidase / Q000096:biosynthesis
chemical_list: D011994:Recombinant Proteins; D014764:Viral Proteins; C062210:bacteriophage T7 RNA polymerase; D012321:DNA-Directed RNA Polymerases; D001616:beta-Galactosidase