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• W2024353610 abstract "Gene delivery into mammalian cells requires the nuclear import of exogenous genes. Since non-viral vectors lack the ability to migrate into the nucleus by themselves, the efficiency of their nuclear import is quite low. Recently, in order to enhance gene transfer, glycosylated carriers were utilized for the nuclear import of plasmid DNA. Although glycofection seems to be potent for enhancing the transfection efficiency and nuclear import of polyplexes, their mechanisms are not clear to date. On the other hand, polyethylenimine (PEI), a synthetic and hyper-branched polyamine, was shown to compact plasmid DNA and to efficiently transfer them into cells both in vitro and vivo. In this work, the mechanism of nuclear import of glycoconjugates consisting of disaccharide- modified PEI/DNA polyplexes is further investigated by using cytoplasmic microinjection. Eight reducing disaccharides (lactose, maltose, isomaltose, mannobiose, mellibiose, gentiobiose, cellobiose, and laminaribiose) were conjugated with branched polyethylenimine (Mw.25 kDa, Aldrich) by reductive amination. The number of sugar moieties linked to PEI was adjusted to a few levels to estimate the potency for enhancing nuclear impot. Disaccharide-modified PEIs were evaluated as non-viral gene carrier to several kinds of cells. Improvement in gene transfer and expression efficiency and the acquirement of cell specificity toward cells could be achieved by the adjustment of number of glycosyl residues. Among compound, 28Lac-PEI with 28% of amino groups substituted with lactosyl residue shows highest transfection efficiency toward not only HepG2 cells which express asialoglycoprotein receptor but also COS-1 and A549 cells which must not express them. Cytoplasmic microinjection of fluorescent labeled polyplex demonstrated that 28Lac-PEI enhanced the import of DNA complex into nucleus, while free PEI, 42Lac-PEI, and 9.3Cel-PEI showed no effect for the nuclear import of plasmid DNA. The nuclear import of 28Lac-PEI/DNA complex occurs through the nuclear pore because it is inhibited by co-microinjection with wheat germ agglutinin, a lectin which blocks the translocation step through the nuclear pore. Furthermore, the nuclear import of 28Lac-PEI/ DNA complex does not use the pathway depending importin-β: co- microinjection of mutant Ran-GTP, inhibitor of the recyclable process of importin- β, resulted in no difference to absence of Ran-GTP. These results demonstrated that lactose-conjugation to cationic non- viral vector increase the transfection efficiency by the enhancement to not only internalization of polyplex but also nuclear import of gene. In addition, nuclear import of glucopolyplex is distinct from importin-β-dependent mechanism." @default.
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• W2024353610 date "2006-01-01" @default.
• W2024353610 modified "2023-10-14" @default.
• W2024353610 title "189. Enhanced Nuclear Import and Transfection Efficiency by Disaccharide-Modified PEIs" @default.
• W2024353610 doi "https://doi.org/10.1016/j.ymthe.2006.08.213" @default.
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