title: Nano-biomimetic carriers are implicated in mechanistic evaluation of intracellular gene delivery.
abstract: Several tissue specific non-viral carriers have been developed for gene delivery purposes. However, the inability to escape endosomes, undermines the efficacy of these carriers. Researchers inspired by HIV and influenza virus, have randomly used Gp41 and H5WYG fusogenic peptides in several gene delivery systems without any rational preference. Here for the first time, we have genetically engineered two Nano-biomimetic carriers composed of either HWYG (HNH) or Gp41 (GNH) that precisely provide identical conditions for the study and evaluation of these fusogenic peptides. The luciferase assay demonstrated a two-fold higher transfection efficiency of HNH compared to GNH. These nanocarriers also displayed equivalent properties in terms of DNA binding ability and DNA protection against serum nucleases and formed similar nanoparticles in terms of surface charge and size. Interestingly, hemolysis and cellular analysis demonstrated both of nanoparticles internalized into cells in similar rate and escaped from endosome with different efficiency. Furthermore, the structural analysis revealed the mechanisms responsible for the superior endosomal escaping capability of H5WYG. In conclusion, this study describes the rationale for using H5WYG peptide to deliver nucleic acids and suggests that using nano-biomimetic carriers to screen different endosomal release peptides, improves gene delivery significantly.
authors.affiliation: Department of Nano biotechnology, Faculty of Biological Sciences, Tarbiat Modares University, Tehran, Iran.
authors.forename: Mohsen
authors.initials: M
authors.lastname: Alipour
authors.affiliation: Department of Biochemistry, Faculty of Biological Sciences, Tarbiat Modares University, Tehran, 14115-175, Iran.
authors.forename: Saman
authors.initials: S
authors.lastname: Hosseinkhani
authors.affiliation: Department of Molecular Biology, Tofigh Daru Co, Tehran, Iran.
authors.forename: Reza
authors.initials: R
authors.lastname: Sheikhnejad
authors.affiliation: Department of Nano biotechnology, Faculty of Biological Sciences, Tarbiat Modares University, Tehran, Iran.
authors.forename: Roya
authors.initials: R
authors.lastname: Cheraghi
mesh_terms: D000595:Amino Acid Sequence; D040761:Biomimetic Materials / Q000737:chemistry*; D016923:Cell Death; D004247:DNA / Q000378:metabolism; D004337:Drug Carriers / Q000737:chemistry; D000067493:Dynamic Light Scattering; D018014:Gene Transfer Techniques; D057809:HEK293 Cells; D006461:Hemolysis; D006801:Humans; D042541:Intracellular Space / Q000378:metabolism*; D008156:Luciferases / Q000378:metabolism; D053758:Nanoparticles / Q000737:chemistry*; D010316:Particle Size; D010455:Peptides / Q000737:chemistry; D044967:Serum / Q000378:metabolism; D014162:Transfection
chemical_list: D004337:Drug Carriers; D010455:Peptides; D004247:DNA; D008156:Luciferases