title: Developing a Novel Gene-Delivery Vector System Using the Recombinant Fusion Protein of Pseudomonas Exotoxin A and Hyperthermophilic Archaeal Histone HPhA.
abstract: 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.
authors.affiliation: Experimental Center of the Functional Subjects, Basic Medical Scientific Research College, China Medical University, Shenyang, Liaoning, P.R.China.
authors.forename: Xin
authors.initials: X
authors.lastname: Deng
authors.affiliation: Institute of Veterinary Medicine, The Academy of Military Medical Sciences of PLA, Changchun, Jilin, P.R. China.
authors.forename: Guoli
authors.initials: G
authors.lastname: Zhang
authors.affiliation: Institute of Veterinary Medicine, The Academy of Military Medical Sciences of PLA, Changchun, Jilin, P.R. China.
authors.forename: Ling
authors.initials: L
authors.lastname: Zhang
authors.affiliation: Key Laboratory for Molecular Enzymology, Jilin University, Changchun, Jilin, P.R.China.
authors.forename: Yan
authors.initials: Y
authors.lastname: Feng
authors.affiliation: Department Biology and Technology of the Agriculture University of Jilin, Changchun, Jilin, P.R.China.
authors.forename: Zehong
authors.initials: Z
authors.lastname: Li
authors.affiliation: Institute of Veterinary Medicine, The Academy of Military Medical Sciences of PLA, Changchun, Jilin, P.R. China.
authors.forename: GuangMou
authors.initials: G
authors.lastname: Wu
authors.affiliation: Institute of Veterinary Medicine, The Academy of Military Medical Sciences of PLA, Changchun, Jilin, P.R. China.
authors.forename: Yuhuan
authors.initials: Y
authors.lastname: Yue
authors.affiliation: Experimental Center of the Functional Subjects, Basic Medical Scientific Research College, China Medical University, Shenyang, Liaoning, P.R.China.|Department of Physiology, China Medical University, Shenyang, Liaoning, P.R.China.
authors.forename: Gensong
authors.initials: G
authors.lastname: Li
authors.affiliation: Department of Physiology, China Medical University, Shenyang, Liaoning, P.R.China.
authors.forename: Yu
authors.initials: Y
authors.lastname: Cao
authors.affiliation: Institute of Veterinary Medicine, The Academy of Military Medical Sciences of PLA, Changchun, Jilin, P.R. China.
authors.forename: Ping
authors.initials: P
authors.lastname: Zhu
mesh_terms: D036002:ADP Ribose Transferases / Q000737:chemistry / Q000235:genetics*; D000818:Animals; D019843:Archaeal Proteins / Q000737:chemistry / Q000235:genetics*; D001427:Bacterial Toxins / Q000737:chemistry / Q000235:genetics*; D016466:CHO Cells; D003412:Cricetulus; D004274:DNA, Recombinant / Q000235:genetics; D005098:Exotoxins / Q000737:chemistry / Q000235:genetics*; D018014:Gene Transfer Techniques; D015316:Genetic Therapy; D005822:Genetic Vectors; D006367:HeLa Cells; D006657:Histones / Q000737:chemistry / Q000235:genetics*; D006801:Humans; D008081:Liposomes; D043902:Pyrococcus horikoshii / Q000235:genetics; D011993:Recombinant Fusion Proteins / Q000737:chemistry / Q000235:genetics; D014162:Transfection / Q000379:methods; D037521:Virulence Factors / Q000737:chemistry / Q000235:genetics*; D000097668:Pseudomonas aeruginosa Exotoxin A
chemical_list: D036002:ADP Ribose Transferases; D019843:Archaeal Proteins; D001427:Bacterial Toxins; D004274:DNA, Recombinant; D005098:Exotoxins; D006657:Histones; D008081:Liposomes; D011993:Recombinant Fusion Proteins; D037521:Virulence Factors; D000097668:Pseudomonas aeruginosa Exotoxin A