Effects of structure of polyamidoamine dendrimer on gene transfer efficiency of the dendrimer conjugate with alpha-cyclodextrin.
To improve gene transfer activity of a new nonviral vector, a polyamidoamine dendrimer (G2) conjugate with alpha-cyclodextrin (alpha-CDE conjugate (G2)), we prepared alpha-CDE conjugates with dendrimer having different generations (G3 and G4), and their gene transfer activities were compared with those of alpha-CDE conjugate (G2) and TransFast, a novel transfection reagent. alpha-CDE conjugates (G2, G3, and G4) formed the complexes with pDNA, changing the zeta-potential and particle size of pDNA complexes and the protection of pDNA from DNase I in a charge ratio-dependent manner, although their differences at higher charge ratios (vector/pDNA) were small. The gene transfer activity of alpha-CDE conjugates (G2, G3, and G4) was higher than that of the corresponding dendrimer alone in NIH3T3 and RAW264.7 cells. Of these CDE conjugates, alpha-CDE conjugate (G3) had a superior gene transfer activity which was comparable to that of TransFast in NIH3T3 cells. The intracellular distribution of pDNA after application of the pDNA complex with alpha-CDE conjugate (G3) to NIH3T3 cells was different from that with dendrimer alone (G3), although the cellular association of pDNA was almost comparable among all vectors. alpha-CDE conjugate (G3) strongly interacted with a fluorescence probe, 2-(p-toluidinyl)-naphthalene-6-sulfonate (TNS), suggesting that the conjugate possesses the inclusion ability with biomembrane constituents such as phospholipids after transfection. These results suggest that alpha-CDE conjugates, particularly the G3 conjugate, could be novel nonviral gene transfer agents.
Kihara
Fumihiro
F
Faculty of Pharmaceutical Sciences, Kumamoto University, 5-1 Oe-honmachi, Kumamoto 862-0973, Japan.
Arima
Hidetoshi
H
Tsutsumi
Toshihito
T
Hirayama
Fumitoshi
F
Uekama
Kaneto
K
D016475:3T3 Cells; D000818:Animals; D001692:Biological Transport; D003505:Cyclodextrins / Q000009:adverse effects / Q000737:chemistry*; D004247:DNA / Q000378:metabolism*; D003850:Deoxyribonuclease I / Q000378:metabolism; D004337:Drug Carriers / Q000009:adverse effects / Q000737:chemistry*; D015870:Gene Expression; D051379:Mice; D010316:Particle Size; D011108:Polymers / Q000009:adverse effects / Q000737:chemistry*; D055672:Static Electricity; D014162:Transfection / Q000379:methods*; D014407:Tumor Cells, Cultured; D047391:alpha-Cyclodextrins
D003505:Cyclodextrins; D004337:Drug Carriers; D011108:Polymers; D047391:alpha-Cyclodextrins; D004247:DNA; D003850:Deoxyribonuclease I; C032613:alpha-cyclodextrin