Efficiency of expression of transfected genes depends on the cell cycle.
Lipofection, a lipid-mediated DNA transfection procedure, was used to transfect synchronized L929 mouse fibroblast cells with a reporter plasmid containing the bacterial chloramphenicol acetyltransferase gene. The efficiency of gene expression was investigated on transfection of cells at different stages of the cell cycle. Our data show that expression of the reporter gene was minimal when transfection was performed in G0-phase and parallel experimental data disproved the possibility that the reduced expression observed was due to differential uptake at different times in the cell cycle. Investigation into the condensation state of the plasmid has shown that the low chloramphenicol acetyltransferase gene expression could be a direct consequence of the packaging of the plasmid into condensed chromatin when transfection occurs in G0-phase. The inactivation of the reporter gene is not reversed by growth of the cells in high serum or by treatment with Trichostatin A, a specific inhibitor of histone deacetylase, suggesting that the inactive chromatin formed in G0-phase cells lacks associated histone acetylase activity. In contrast, the high activity seen when cells in S-phase are transfected is enhanced even further by treatment with Trichostatin A.
Marenzi
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S
Department of Biological Science A. Rossi-Fanelli, University of Rome La Sapienza, Italy.
Adams
R L
RL
Zardo
G
G
Lenti
L
L
Reale
A
A
Caiafa
P
P
D000123:Acetyltransferases / Q000378:metabolism; D000818:Animals; D002453:Cell Cycle; D002460:Cell Line; D015500:Chloramphenicol O-Acetyltransferase / Q000235:genetics* / Q000378:metabolism*; D002843:Chromatin / Q000737:chemistry / Q000378:metabolism; D004791:Enzyme Inhibitors / Q000494:pharmacology; D005347:Fibroblasts; D015870:Gene Expression; D017930:Genes, Reporter; D051548:Histone Acetyltransferases; D056572:Histone Deacetylase Inhibitors; D006877:Hydroxamic Acids / Q000494:pharmacology; D051379:Mice; D010957:Plasmids / Q000235:genetics; D029701:Saccharomyces cerevisiae Proteins; D014162:Transfection / Q000379:methods*
D002843:Chromatin; D004791:Enzyme Inhibitors; D056572:Histone Deacetylase Inhibitors; D006877:Hydroxamic Acids; D029701:Saccharomyces cerevisiae Proteins; C012589:trichostatin A; D000123:Acetyltransferases; D015500:Chloramphenicol O-Acetyltransferase; D051548:Histone Acetyltransferases