Expression behavior of high-pressure-compacted plasmid DNA in mammalian cell.
We have been developing a novel compaction method of plasmid DNA using high pressure technology, and previously found that the size of the plasmid DNA was decreased with increasing the pressurizing-strength and time. In the present study, we investigated the tertiary structural change and the expression behavior of the pressure-compacted plasmid DNA in cell in vitro. When the pressure-compacted plasmid DNA was reacted with restriction enzyme (EcoRI), a large amount of the EcoRI was required to cleave the pressure-compacted plasmid DNA than the non-pressurized plasmid DNA, suggesting that the structural change of plasmid DNA was induced by the pressurization. The expression of the pressure-compacted plasmid DNA injected into cells using microinjection method was analyzed. It was clear that the pressure-compacted plasmid DNA could effectively delay gene expression, suggesting that the controlled compaction of plasmid DNA by high pressurization would be regulate the transgene expression.
Kimura
Tsuyoshi
T
Institute of Biomaterials and Bioengineering, Tokyo Medical and Dental University, Tokyo 101-0062, Japan.
Konno
Hokuto
H
Fujisato
Toshiya
T
Kishida
Akio
A
D004247:DNA / Q000737:chemistry* / Q000378:metabolism; D015246:Deoxyribonuclease EcoRI / Q000378:metabolism; D015870:Gene Expression; D006367:HeLa Cells; D006801:Humans; D006874:Hydrostatic Pressure; D009690:Nucleic Acid Conformation; D010957:Plasmids / Q000235:genetics*; D019076:Transgenes
D004247:DNA; D015246:Deoxyribonuclease EcoRI