Evaluation of polyether-polyethyleneimine graft copolymers as gene transfer agents
Evaluation of polyether-polyethyleneimine graft copolymers as gene transfer agents
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DOI：
10.1038/sj.gt.3301052
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发表时间：
2000-01-01
期刊：
影响因子：
5.1
通讯作者：
Kabanov, AV
中科院分区：
文献类型：
--
作者：
Nguyen, HK;Lemieux, P;Kabanov, AV
阳离子共聚物由连接到非离子聚合物的阳离子组成，被评价为非病毒基因传递系统。已知这些共聚物可与DNA产生可溶性复合物，但只有少数研究表征了这些复合物的转染活性。本文报道了一系列阳离子共聚物的合成和表征，这些阳离子共聚物是由聚乙烯亚胺（PEI）与非离子聚醚、聚环氧乙烷（PEO）或Pluronic 123 （P123）接枝而成的。PEO-PEI偶联物在PN的分子质量（2 kDa和25 kDa）和PN被PEG修饰的程度上有所不同。所有这些缀合物在与质粒混合后形成复合物，质粒在水分散中稳定数天。根据复合物的组成，在这些体系中形成的颗粒的大小从70到200纳米不等。然而，在体外转染实验中，这些系统的转染活性远低于PN （25 kDa）或sufect，使用荧光素酶报告基因在Cos-7细胞中表达作为模型系统，相比之下，P123与PEI （2 kDa）混合自由P123 （9:1(wt)）的偶联物与DNA （110 nm）形成小而稳定的复合物，在体外表现出高转染活性。此外，小鼠静脉注射该复合物24 h后，在脾脏、心脏、肺和肝脏中观察到基因表达。与1,2-bis（油酰氧基）-（三甲基氨酰基）丙烷：胆固醇（DOTAP:Chol）和PN （25 kDa）转染系统相比，P123-PEI系统在这些器官之间的基因表达分布更为均匀，可以显著改善肝脏中的基因表达。
Cationic copolymers consisting of polycations linked to nonionic polymers are evaluated as non-viral gene delivery systems. These copolymers are known to produce soluble complexes with DNA, but only a few studies have characterized the transfection activity of these complexes. This work reports the synthesis and characterization of a series of cationic copolymers obtained by grafting the polyethyleneimine (PEI) with non-ionic polyethers, poly (ethylene oxide) (PEO) or Pluronic 123 (P123). The PEO-PEI conjugates differ in the molecular mass of PN (2 kDa and 25 kDa) and the degree of modification of PN with PEG. All of these conjugates form complexes upon mixing with plasmids, which are stable in aqueous dispersion for several days. The sizes of the particles formed in these systems vary from 70 to 200 nm depending on the composition of the complex. However, transfection activity of these systems is much lower than that of PN (25 kDa) or Superfect as assessed in in vitro transfection experiments utilizing a luciferase reporter expression in Cos-7 cells as a model system, in contrast, conjugate of P123 with PEI (2 kDa) mixed with free P123 (9:1(wt)) forms small and stable complexes with DNA (110 nm) that exhibit high transfection activity in vitro. Furthermore, gene expression is observed in spleen, heart, lungs and liver 24 h after i.v. injection of this complex in mice. Compared to 1,2-bis(oleoyloxy)-(trimethylammonio) propane:cholesterol (DOTAP:Chol) and PN (25 kDa) transfection systems, the P123-PEI system reveals a more uniform distribution of gene expression between these organs, allowing a significant improvement of gene expression in liver.