Influence of promoter choice and trichostatin A treatment on expression of baculovirus delivered genes in mammalian cells.
The expression of recombinant proteins following transduction of CHO cells with recombinant baculoviruses containing a mammalian expression cassette with the CMV-promoter is enhanced by the addition of trichostatin A (TSA), a specific histone deacetylase inhibitor. To further investigate the effect of TSA treatment on protein production following BacMam transduction, viruses containing various viral promoters (SV40, CMV, and RSV) and one cellular promoter (human ubiquitin C) were compared with regard to expression level of a gfp-luciferase fusion protein following transduction of CHO, COS-1, and HEK293 cells. The overall effect on expression appears to be cell specific, indicating that different mechanisms are active within different cell lines. Further, COS cells transfected with naked viral DNA, plasmids, and baculovirus particles were compared in regard to TSA treatment. The increase in reporter gene expression observed following BacMam transduction and TSA treatment were greater than those for transfection of either naked viral DNA or plasmid DNA.
Spenger
Alexandra
A
Institute of Applied Microbiology, University of Natural Resources and Applied Life Sciences, Vienna, Muthgasse 18, A-1190 Vienna, Austria.
Ernst
Wolfgang
W
Condreay
J Patrick
JP
Kost
Thomas A
TA
Grabherr
Reingard
R
D000818:Animals; D016367:Baculoviridae / Q000187:drug effects / Q000235:genetics*; D016466:CHO Cells; D019556:COS Cells; D006224:Cricetinae; D003412:Cricetulus; D005798:Genes, Bacterial; D006877:Hydroxamic Acids / Q000494:pharmacology*; D011401:Promoter Regions, Genetic; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism
D006877:Hydroxamic Acids; D011993:Recombinant Fusion Proteins; C012589:trichostatin A