A lipoplex （i.e., pDNA#1/lipid complex and transfection reagent for pDNA delivery） containing galactosylceramide （GalCer） and an amidine-bearing lipid （TRX） was examined whether the bound pDNA was specifically i...A lipoplex （i.e., pDNA#1/lipid complex and transfection reagent for pDNA delivery） containing galactosylceramide （GalCer） and an amidine-bearing lipid （TRX） was examined whether the bound pDNA was specifically ingested by hepatocyte via asialoglycoprotein receptor （ASGPR） and then expressed protein. Gel electrophoresis and small-angle X-ray scattering （SAXS） confirmed that the TRX-GalCer liposome#2 complexed with pDNA and the resultant lipoplex took a hexagonally packed inverted cylinder structure when the GalCer composition was less than 20 wt.% of the total lipid. When the lipoplex carrying pGL3 （luciferase-cording pDNA） was administrated to HepG2, the luciferase activity was increased with increasing the GalCer composition until it reached 3 wt.% and then decreased upon further addition of GalCer. When we added galactose itself as a competitor, the luciferase activity was decreased, while glucose did not show such decrease, suggesting that HepG2 ingested the lipoplex via ASGPR-mediated endocytosis. This paper indicated that the hexagonally packed inverted cylinder structures of lipoplex may not always provide excellent transfection and presented a possibility that the TRX lipoplex#3 can obtain a cellulartargeting ability through the receptors for oligosaccharide.展开更多
基金SORST of JST and a Grant-in-Aid for Scientific Research(No.16350068 and 16655048)SAXS was performed at SPring-8 BL40B2(No.2006A1510).
文摘A lipoplex （i.e., pDNA#1/lipid complex and transfection reagent for pDNA delivery） containing galactosylceramide （GalCer） and an amidine-bearing lipid （TRX） was examined whether the bound pDNA was specifically ingested by hepatocyte via asialoglycoprotein receptor （ASGPR） and then expressed protein. Gel electrophoresis and small-angle X-ray scattering （SAXS） confirmed that the TRX-GalCer liposome#2 complexed with pDNA and the resultant lipoplex took a hexagonally packed inverted cylinder structure when the GalCer composition was less than 20 wt.% of the total lipid. When the lipoplex carrying pGL3 （luciferase-cording pDNA） was administrated to HepG2, the luciferase activity was increased with increasing the GalCer composition until it reached 3 wt.% and then decreased upon further addition of GalCer. When we added galactose itself as a competitor, the luciferase activity was decreased, while glucose did not show such decrease, suggesting that HepG2 ingested the lipoplex via ASGPR-mediated endocytosis. This paper indicated that the hexagonally packed inverted cylinder structures of lipoplex may not always provide excellent transfection and presented a possibility that the TRX lipoplex#3 can obtain a cellulartargeting ability through the receptors for oligosaccharide.
