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基于蛋白质作为核心骨架构建一种新型非病毒基因递送载体。

The construction of a novel kind of non-viral gene delivery vector based on protein as core backbone.

作者信息

Li D, Kong Y, Yu H, Lehtinen A, Huang H, Shen F, Min L, Zhou J, Tang G, Wang Q

机构信息

Department of Oncology, Sir Run Run Shaw Hospital affiliated to School of Medicine, Zhejiang University, Hangzhou 310003, China.

Institute of Chemical Biology and Pharmaceutical Chemistry, Zhejiang University, Hangzhou 310028, China.

出版信息

Vox Sang. 2008 Apr;94(3):234-241. doi: 10.1111/j.1423-0410.2007.01025.x. Epub 2007 Dec 19.

Abstract

BACKGROUND AND OBJECTIVES

A novel kind of non-viral gene delivery vector based on transferrin (Tf) as the core component was constructed with high transfection efficiency and low toxicity.

MATERIALS AND METHODS

The synthesis vector of Tf-PEI600 was confirmed by different physicochemical methods, including (1)H nuclear magnetic resonance, gel permeation chromatography, X-ray and thermogravimetric analysis. The cytotoxicity and gene delivery efficiency of the synthesized vector were verified by in vitro experiments.

RESULTS

The agarose gel electrophoresis assay indicated that the novel copolymer Tf-PEI600 could efficiently condense plasmid DNA and the condensed nanoparticles exhibited a spherical shape. As the weight ratio of Tf-PEI600 to DNA reached 15.0, the particle size (about 200 nm) and the zeta potential (about 20 mV) of the nanoparticles became optimal for gene delivery. The methylthiazolyl tetrazolium (MTT) assay showed the cytotoxicity of Tf-PEI600 to be similar to that of PEI600 and much lower than that of PEI25kDa. In gene-delivery experiments with COS-7 cells and HepG2 cells, the Tf-PEI600 showed about a 30- to 53-fold higher efficiency than PEI600 and nearly equal to that of PEI25kDa.

CONCLUSIONS

These data suggest that Tf-PEI600, with the advantages of low toxicity and high gene-delivery efficiency, might have great prospects in the practice of gene delivery. The core-shell structure of Tf-PEI600 also provided a novel strategy for the construction of non-viral gene delivery vectors.

摘要

背景与目的

构建了一种以转铁蛋白(Tf)为核心成分的新型非病毒基因递送载体,其具有高转染效率和低毒性。

材料与方法

通过不同的物理化学方法对Tf-PEI600合成载体进行了确认,包括氢核磁共振(¹H NMR)、凝胶渗透色谱法、X射线和热重分析。通过体外实验验证了合成载体的细胞毒性和基因递送效率。

结果

琼脂糖凝胶电泳分析表明,新型共聚物Tf-PEI600能够有效地浓缩质粒DNA,且浓缩后的纳米颗粒呈球形。当Tf-PEI600与DNA的重量比达到15.0时,纳米颗粒的粒径(约200 nm)和zeta电位(约20 mV)对于基因递送而言变得最为理想。甲基噻唑基四氮唑(MTT)分析显示,Tf-PEI600的细胞毒性与PEI600相似,且远低于PEI25kDa。在COS-7细胞和HepG2细胞的基因递送实验中,Tf-PEI600的效率比PEI600高约30至53倍,且几乎与PEI25kDa相当。

结论

这些数据表明,Tf-PEI600具有低毒性和高基因递送效率的优点,在基因递送实践中可能具有广阔前景。Tf-PEI600的核壳结构也为非病毒基因递送载体的构建提供了一种新策略。

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