Enhancement of gene expression by polyamidoamine dendrimer conjugates with alpha-, beta-, and gamma-cyclodextrins.
To improve the transfection efficiency of nonviral vector, we synthesized the starburst polyamidoamine dendrimer conjugates with alpha-, beta-, and gamma-cyclodextrins (CDE conjugates), expecting the synergistic effect of dendrimer and cyclodextrins (CyDs). The (1)H NMR spectroscopic data indicated that alpha-, beta-, and gamma-CyDs are covalently bound to dendrimer in a molar ratio of 1:1. The agarose gel electrophoretic studies revealed that CDE conjugates formed the complexes with plasmid DNA (pDNA) and protected the degradation of pDNA by DNase I in the same manner as dendrimer. CDE conjugates showed a potent luciferase gene expression, especially in the dendrimer conjugate with alpha-CyD (alpha-CDE conjugate) which provided the greatest transfection activity (approximately 100 times higher than those of dendrimer alone and of the physical mixture of dendrimer and alpha-CyD) in NIH3T3 and RAW264.7 cells. In addition, the gene transfer activity of alpha-CDE conjugate was superior to that of Lipofectin. The enhancing gene transfer effect of alpha-CDE conjugate may be attributable to not only increasing the cellular association, but also changing the intracellular trafficking of pDNA. These findings suggest that alpha-CDE conjugate could be a new preferable nonviral vector of pDNA.
Arima
H
H
Faculty of Pharmaceutical Sciences, Kumamoto University, 5-1 Oe-honmachi, Kumamoto 862-0973, Japan.
Kihara
F
F
Hirayama
F
F
Uekama
K
K
D016475:3T3 Cells / Q000378:metabolism; D000818:Animals; D001665:Binding Sites / Q000502:physiology; D003505:Cyclodextrins / Q000737:chemistry / Q000494:pharmacology*; D004247:DNA / Q000737:chemistry / Q000378:metabolism*; D003850:Deoxyribonuclease I / Q000378:metabolism*; D004357:Drug Synergism; D015870:Gene Expression / Q000187:drug effects*; D018014:Gene Transfer Techniques; D005822:Genetic Vectors / Q000235:genetics; D051379:Mice; D018613:Microscopy, Confocal; D010714:Phosphatidylethanolamines / Q000494:pharmacology; D010957:Plasmids / Q000737:chemistry; D011108:Polymers / Q000494:pharmacology*; D014162:Transfection
D003505:Cyclodextrins; D010714:Phosphatidylethanolamines; D011108:Polymers; C020888:1,2-dielaidoylphosphatidylethanolamine; D004247:DNA; D003850:Deoxyribonuclease I