Wang Zu-Yong, Zhao Yang, Ren Lei, Jin Li-Hua, Sun Li-Ping, Yin Pei, Zhang Ya-Fei, Zhang Qi-Qing
Research Center of Biomedical Engineering, Department of Biomaterials, College of Materials, Xiamen University, Xiamen 361005, People's Republic of China.
Nanotechnology. 2008 Nov 5;19(44):445103. doi: 10.1088/0957-4484/19/44/445103. Epub 2008 Sep 26.
In principle, the technique of gene delivery involves taking complete or parts of genes that can code specific messages and delivering them to selected cells in the body. Such a transfer of plasmid DNA into mammalian cells has posed major challenges for gene therapy. A series of gelatin-siloxane nanoparticles (GS NPs) with controlled size and surface charge were synthesized through a two-step sol-gel process. In order to increase the efficiency of cellular uptake, HIV-derived Tat peptide was further grafted to GS NPs. In vitro co-location and endocytosis inhibition experiments suggested that the as-synthesized TG NPs may enter HeLa cells via a combined pathway of lipid-raft- and receptor-dependent endocytosis, and only cause little cell damage. Moreover, this study shows the encapsulation of a plasmid DNA in TG NPs to be obtained as a non-viral gene vector. This kind of encapsulation provides complete protection to the plasmid DNA from the external DNase and serum environment, and generates the hope that the resulting formulation can be developed into a potential vector for effective gene delivery. In order to check this potential, the reporter gene pSVβ-gal was encapsulated, and in vitro transfection efficiency of this system was found to be nearly 130% compared to the commercially available transfection reagent Lipofectamine™.
原则上,基因传递技术涉及获取能够编码特定信息的完整基因或部分基因,并将其传递至体内特定细胞。将质粒DNA转移至哺乳动物细胞对基因治疗构成了重大挑战。通过两步溶胶 - 凝胶法合成了一系列尺寸和表面电荷可控的明胶 - 硅氧烷纳米颗粒(GS NPs)。为了提高细胞摄取效率,将源自HIV的Tat肽进一步接枝到GS NPs上。体外共定位和内吞抑制实验表明,合成的TG NPs可能通过脂筏和受体依赖性内吞的联合途径进入HeLa细胞,并且仅造成轻微的细胞损伤。此外,本研究表明将质粒DNA封装在TG NPs中可作为非病毒基因载体。这种封装为质粒DNA提供了对外部DNase和血清环境的完全保护,并带来了希望,即所得制剂可开发成用于有效基因传递的潜在载体。为了检验这种潜力,将报告基因pSVβ - gal进行封装,并且发现该系统的体外转染效率与市售转染试剂Lipofectamine™相比接近130%。