Expression profile of plasmid DNA by spermine derivatives of pullulan with different extents of spermine introduced
Expression profile of plasmid DNA by spermine derivatives of pullulan with different extents of spermine introduced
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DOI：
10.1016/j.jconrel.2007.01.005
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发表时间：
2007-04-23
影响因子：
10.8
通讯作者：
Tabata, Yasuhiko
中科院分区：
文献类型：
--
作者：
Jo, Jun-ichiro;Kai, Tomonori;Tabata, Yasuhiko
本研究的目的是利用普鲁兰多糖制备一种新型的基因载体，并评价其用于基因转染的可行性。以分子量为22，800、47，300和112，000的普鲁兰多糖为原料，通过化学方法引入不同量的精胺，制备了不同精胺含量的阳离子普鲁兰多糖衍生物。将每种阳离子化普鲁兰衍生物与质粒DNA以各种比例复合，并应用于HepG2细胞进行体外基因转染。基因表达水平取决于阳离子化普鲁兰衍生物引入精胺的百分比和普鲁兰的分子量。然而，当以普鲁兰多糖衍生物与质粒DNA的络合摩尔比进行比较时，表达水平在10（2）的比率附近变得最大，而与普鲁兰多糖分子量无关。用去唾液酸糖蛋白受体配体的去唾液酸胎球蛋白预处理细胞降低了复合物的基因表达水平。阳离子化普鲁兰多糖衍生物具有良好的理化性质，是一种很有前途的非病毒载体，可促进受体介导的质粒DNA内化，从而提高表达水平。(c)2007 Elsevier B.V.保留所有权利。
The objective of this study is to prepare a novel gene carrier from pullulan, a polysaccharide with an inherent affinity for the liver, and evaluate the feasibility in gene transfection. Various amounts of spermine were chemically introduced into pullulan with molecular weights of 22,800, 47,300, and 112,000 to prepare cationized pullulan derivatives with different percentages of spermine introduced. Each cationized pullulan derivative was complexed with a plasmid DNA at various ratios and applied to HepG2 cells for in vitro gene transfection. The level of gene expression depended on the percent spermine introduced of cationized pullulan derivatives and the molecular weight of pullulan. However, when compared at the complexation molar ratio of pullulan derivative to the plasmid DNA, the expression level became maximum around the ratio of 10(2), irrespective of the pullulan molecular weight. Pre-treatment of cells with asialofetuin of asialoglycoprotein receptor ligand decreased the level of gene expression by the complexes. The cationized pullulan derivative with an appropriate physicochemical character is a promising non-viral carrier which promotes the receptor-mediated internalization of plasmid DNA and consequently enhances the expression level. (c) 2007 Elsevier B.V. All rights reserved.