Developing a Novel Gene-Delivery Vector System Using the Recombinant Fusion Protein of Pseudomonas Exotoxin A and Hyperthermophilic Archaeal Histone HPhA.
Developing a Novel Gene-Delivery Vector System Using the Recombinant Fusion Protein of Pseudomonas Exotoxin A and Hyperthermophilic Archaeal Histone HPhA.
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利用假单胞菌外毒素 A 和超嗜热古菌组蛋白 HPhA 的重组融合蛋白开发新型基因递送载体系统
DOI：
10.1371/journal.pone.0142558
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发表时间：
2015
期刊：
影响因子：
3.7
通讯作者：
Zhu P
中科院分区：
文献类型：
--
作者：
Deng X;Zhang G;Zhang L;Feng Y;Li Z;Wu G;Yue Y;Li G;Cao Y;Zhu P
非病毒基因传递系统具有许多优点，是基因治疗的发展方向。这种方法的一个固有障碍是通过内吞作用摄取到囊泡隔室中。受体介导的基因传递方法有望克服这一障碍。在这项研究中，我们开发了一个受体介导的基因传递系统的基础上的组合的假单胞菌外毒素A（PE），它具有受体结合和膜转位结构域，和超嗜热古菌组蛋白（HPhA），它具有DNA结合能力。首先，我们构建并表达了rPE-HPhA融合蛋白。然后我们检测了rPE-HPhA的细胞毒性和DNA结合能力。我们进一步评估了通过rPE-HPhA系统将pEGF-C1质粒DNA转染至CHO细胞的效率，并与阳离子脂质体方法进行了比较。结果表明，rPE-HPhA的转染效率高于阳离子脂质体。此外，rPE-HPhA基因递送系统对DNA序列、拓扑结构或靶细胞类型是非特异性的。因此，rPE-HPhA系统可用于将感兴趣的基因递送到哺乳动物细胞中，并且具有应用于基因治疗的巨大潜力。
Non-viral gene delivery system with many advantages has a great potential for the future of gene therapy. One inherent obstacle of such approach is the uptake by endocytosis into vesicular compartments. Receptor-mediated gene delivery method holds promise to overcome this obstacle. In this study, we developed a receptor-mediated gene delivery system based on a combination of the Pseudomonas exotoxin A (PE), which has a receptor binding and membrane translocation domain, and the hyperthermophilic archaeal histone (HPhA), which has the DNA binding ability. First, we constructed and expressed the rPE-HPhA fusion protein. We then examined the cytotoxicity and the DNA binding ability of rPE-HPhA. We further assessed the efficiency of transfection of the pEGF-C1 plasmid DNA to CHO cells by the rPE-HPhA system, in comparison to the cationic liposome method. The results showed that the transfection efficiency of rPE-HPhA was higher than that of cationic liposomes. In addition, the rPE-HPhA gene delivery system is non-specific to DNA sequence, topology or targeted cell type. Thus, the rPE-HPhA system can be used for delivering genes of interest into mammalian cells and has great potential to be applied for gene therapy.