title: Design, engineering and preparation of a multi-domain fusion vector for gene delivery.
abstract: Peptide based gene carriers are among the most promising non-viral vectors for gene delivery to eukaryotic cells. We have engineered a new fusion peptide using recombinant technology with the purpose of overcoming the cell barriers to gene delivery. A His- tagged multi-domain peptide was expressed in Escherichia coli BL21 (DE3) pLysS and purified using Ni-NTA resin. The fusion peptide is composed of two repeats of truncated histone H1 peptide to condense pDNA, a fusogenic peptide to disrupt endosome membranes and a nuclear localization signal to enhance translocation of pDNA towards nucleus. The results demonstrated that the vector can effectively condense plasmid DNA into nanoparticles with average sizes of 200 nm. The fusogenic peptide in the vector structure also showed membrane disruptive effect in the endosomal pH. Overall, the transfection efficiency of the vector demonstrated that it holds great promise as a nontoxic and effective non-viral gene carrier.
authors.affiliation: Department of Nanobiotechnology, Faculty of Biological Sciences, Tarbiat Modares University, Tehran, Iran.
authors.forename: Faranak
authors.initials: F
authors.lastname: Sadeghian
authors.forename: Saman
authors.initials: S
authors.lastname: Hosseinkhani
authors.forename: Abdolali
authors.initials: A
authors.lastname: Alizadeh
authors.forename: Arash
authors.initials: A
authors.lastname: Hatefi
mesh_terms: D000818:Animals; D016466:CHO Cells; D002738:Chloroquine / Q000494:pharmacology; D003001:Cloning, Molecular; D006224:Cricetinae; D003412:Cricetulus; D004247:DNA / Q000097:blood; D003851:Deoxyribonucleases / Q000737:chemistry; D016503:Drug Delivery Systems; D015195:Drug Design; D004563:Electrochemistry; D004926:Escherichia coli / Q000235:genetics; D018014:Gene Transfer Techniques; D005818:Genetic Engineering; D005822:Genetic Vectors / Q000008:administration & dosage*; D006461:Hemolysis / Q000187:drug effects; D008027:Light; D010316:Particle Size; D010957:Plasmids / Q000008:administration & dosage / Q000235:genetics; D011994:Recombinant Proteins / Q000235:genetics; D012542:Scattering, Radiation; D012685:Sepharose / Q000737:chemistry; D014162:Transfection
chemical_list: D011994:Recombinant Proteins; D002738:Chloroquine; D004247:DNA; D012685:Sepharose; D003851:Deoxyribonucleases