Transient and stable gene expression in mammalian cells transduced with a recombinant baculovirus vector.
Recombinant baculoviruses can serve as gene-transfer vehicles for transient expression of recombinant proteins in a wide range of mammalian cell types. Furthermore, by inclusion of a dominant selectable marker in the viral vector, cell lines can be derived that stably express recombinant genes. A virus was constructed containing two expression cassettes controlled by constitutive mammalian promoters: the cytomegalovirus immediate early promoter/enhancer directing expression of green fluorescent protein and the simian virus 40 (SV40) early promoter controlling neomycin phosphotransferase II. Using this virus, efficient gene delivery and expression was observed and measured in numerous cell types of human, primate, and rodent origin. In addition to commonly used transformed cell lines such as HeLa, CHO, Cos-7, and 293, this list includes primary human keratinocytes and bone marrow fibroblasts. In all cases, addition of butyrate or trichostatin A (a selective histone deacetylase inhibitor) to transduced cells markedly enhanced the levels of reporter protein expression observed. When transduced cells are put under selection with the antibiotic G418, cell lines can be obtained at high frequency that stably maintain the expression cassettes of the vector DNA and exhibit stable, high-level expression of the reporter gene. Stably transduced derivatives have been selected from a substantial number of different cell types, suggesting that stable lines can be derived from any cell type that exhibits transient expression.
Condreay
J P
JP
Glaxo Wellcome Research Institute, P.O. Box 13398, Research Triangle Park, NC 27709, USA. jpc37987@glaxowellcome.com
Witherspoon
S M
SM
Clay
W C
WC
Kost
T A
TA
D000818:Animals; D016367:Baculoviridae / Q000235:genetics* / Q000648:ultrastructure; D016466:CHO Cells; D019556:COS Cells; D002478:Cells, Cultured; D006224:Cricetinae; D003587:Cytomegalovirus / Q000235:genetics; D015870:Gene Expression; D005822:Genetic Vectors; D049452:Green Fluorescent Proteins; D006367:HeLa Cells; D056572:Histone Deacetylase Inhibitors; D006801:Humans; D006877:Hydroxamic Acids / Q000494:pharmacology; D008164:Luminescent Proteins / Q000096:biosynthesis / Q000235:genetics; D012377:Rodentia; D013539:Simian virus 40 / Q000235:genetics; D014170:Transformation, Genetic
D056572:Histone Deacetylase Inhibitors; D006877:Hydroxamic Acids; D008164:Luminescent Proteins; D049452:Green Fluorescent Proteins; C012589:trichostatin A