Transition from a normal to inverted cylinder for an amidine-bearing lipid/pDNA complex and its excellent transfection.
A cationic lipid (TRX) having an amidine headgroup was synthesized, and a lipoplex (i.e., plasmid DNA+lipid complex) was prepared from the mixture of TRX and two other neutral colipids. Small-angle X-ray scattering showed that the addition of DNA induced a structural transition from normal to inverted hexagonally packed cylinders. Transmission electron microscopy showed a threadlike micelle was transformed to a spherical micelle about 50-200 nm in diameter by adding DNA. A combination of these results leads to the conclusion that the complexed DNA is hexagonally packed (or condensed) into spherical aggregates. The size and morphology are believed suitable for endocytosis uptake or vesicle fusion. The complex made from pDNA (pEGFP-C1) and TRX was transfected to Hep G2. Flow cytometry and confocal microscopy showed that the present system expressed green fluorescence protein (GFP) more than a conventional transfection reagent. Additionally, TRX was less cytotoxic than other transfection reagents. This paper presents the attractive possibility of the amidine group for a transfection device.
Koiwai
Kazunori
K
Terumo Corporation R & D Center, 1500 Inokuchi, Nakai-machi, Ashigarakami-gun Kanagawa, 259-0151, Japan.
Tokuhisa
Kenji
K
Karinaga
Ryouji
R
Kudo
Yasuya
Y
Kusuki
Shota
S
Takeda
Yoichi
Y
Sakurai
Kazuo
K
D000578:Amidines; D002412:Cations; D045744:Cell Line, Tumor; D049452:Green Fluorescent Proteins / Q000235:genetics; D006801:Humans; D008055:Lipids / Q000737:chemistry*; D008823:Micelles; D010316:Particle Size; D044367:Phase Transition; D010957:Plasmids / Q000737:chemistry*; D014162:Transfection / Q000379:methods*
D000578:Amidines; D002412:Cations; D008055:Lipids; D008823:Micelles; D049452:Green Fluorescent Proteins