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基于聚(2-乙基-2-恶唑啉)与线性聚乙烯亚胺的二嵌段共聚物的非病毒基因载体

Nonviral gene carriers based on diblock copolymers of poly(2-ethyl-2-oxazoline) and linear polyethylenimine.

作者信息

Hsiue Ging-Ho, Chiang Hou-Zen, Wang Chau-Hui, Juang Tzuoh-Min

机构信息

Department of Chemical Engineering, National Tsing Hua University, Hsinchu, 300 Taiwan, ROC. ghhsiue@ che.nthu.edu.tw

出版信息

Bioconjug Chem. 2006 May-Jun;17(3):781-6. doi: 10.1021/bc050317u.

Abstract

Diblock copolymers that consist of poly(2-ethyl-2-oxazoline) (PEOz) and linear polyethylenimine (LPEI) were prepared for use as nonviral gene carriers. The PEOz-b-LPEI copolymers were synthesized by coupling PEOz with LPEI in a thiol-disulfide exchange reaction between the sulfhydryl and pyridyl disulfide terminal groups. A polymer/DNA weight ratio (P/D) of over 12 was required to enable PEOz-b-LPEI to condense DNA completely. The DNA-condensing capability of the diblock copolymers was increased with increasing the hydrolytic degrees of the LPEI segment. The PEOz-b-LPEI polyplexes were stable in 150 mM NaCl aqueous solution and had a mean diameter around 190 nm, whereas BPEI and LPEI polyplexes formed large aggregates in the range 300-500 nm. In addition, these polyplexes exhibited the sensitivity to solution pH and were dissociated in the acidic buffers (pH < or = 5.5). The results of in vitro cell viability and luciferase assay indicated that PEOz-b-LPEI showed not only low cytotoxicity but also high transfection efficiency in gene expression.

摘要

制备了由聚(2-乙基-2-恶唑啉)(PEOz)和线性聚乙烯亚胺(LPEI)组成的双嵌段共聚物,用作非病毒基因载体。通过在巯基和吡啶二硫端基之间的硫醇-二硫交换反应中使PEOz与LPEI偶联来合成PEOz-b-LPEI共聚物。需要聚合物/DNA重量比(P/D)超过12才能使PEOz-b-LPEI完全凝聚DNA。随着LPEI链段水解度的增加,双嵌段共聚物的DNA凝聚能力增强。PEOz-b-LPEI多聚体在150 mM NaCl水溶液中稳定,平均直径约为190 nm,而BPEI和LPEI多聚体形成300-500 nm范围内的大聚集体。此外,这些多聚体对溶液pH敏感,在酸性缓冲液(pH≤5.5)中会解离。体外细胞活力和荧光素酶测定结果表明,PEOz-b-LPEI不仅具有低细胞毒性,而且在基因表达中具有高转染效率。

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