Developing a Novel Gene-Delivery Vector System Using the Recombinant Fusion Protein of Pseudomonas Exotoxin A and Hyperthermophilic Archaeal Histone HPhA

• Published in: PloS one Vol. 10; no. 11; p. e0142558
• Main Authors: Deng, Xin, Zhang, Guoli, Zhang, Ling, Feng, Yan, Li, Zehong, Wu, GuangMou, Yue, Yuhuan, Li, Gensong, Cao, Yu, Zhu, Ping
• Format: Journal Article
• Language: English
• Published: United States Public Library of Science 10.11.2015; Public Library of Science (PLoS)
• Subjects: ADP Ribose Transferases - chemistry; ADP Ribose Transferases - genetics; Animals; Archaeal Proteins - chemistry; Archaeal Proteins - genetics; Bacterial Toxins - chemistry; Bacterial Toxins - genetics; Binding; Cancer; Cations; CHO Cells; Cloning; Compartments; Cricetulus; Cytotoxicity; Deoxyribonucleic acid; DNA; DNA, Recombinant - genetics; E coli; Efficiency; Endocytosis; Exotoxin A; Exotoxins - chemistry; Exotoxins - genetics; Fusion protein; Fusion proteins; Gene therapy; Gene transfer; Gene Transfer Techniques; Genetic aspects; Genetic Therapy; Genetic Vectors; HeLa Cells; Histones - chemistry; Histones - genetics; Humans; Innovations; Liposomes; Mammalian cells; Nucleotide sequence; Physiology; Plasmid DNA; Plasmids; Properties; Proteins; Pseudomonas; Pseudomonas aeruginosa; Pseudomonas aeruginosa Exotoxin A; Pyrococcus horikoshii; Pyrococcus horikoshii - genetics; Recombinant Fusion Proteins - chemistry; Recombinant Fusion Proteins - genetics; Topology; Toxicity; Transfection; Transfection - methods; Translocation; Vectors (Biology); Veterinary medicine; Virulence Factors - chemistry; Virulence Factors - genetics
• ISSN: 1932-6203; 1932-6203
• DOI: 10.1371/journal.pone.0142558

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.