title: Quaternary ammonium salt containing soybean oil: an efficient nanosize gene delivery carrier for halophile green microalgal transformation.
abstract: Dunaliella salina, a halophile green microalga, is considered a robust photobioreactor and a remarkable cost beneficial system for the production of therapeutic recombinant proteins. In this study, with low overall cost, a proper cationic lipid was synthesized from renewable soybean oil as an efficient gene delivery carrier for D. salina cells to create appropriate protein-producing transformed cell lines. To obtain an effective carrier, quaternary ammonium salt containing soybean oil (QASSO) was synthesized through the ring opening reaction of the epoxy groups of epoxidized soybean oil with diethylamine. QASSO was characterized using nuclear magnetic resonance and Fourier-transform infrared instruments. QASSO was used to prepare nanolipoplex construct using plasmid DNA molecules containing green fluorescent protein (GFP) as reporter gene. These nanolipoplexes (QASSO-pGFP, N/P=3) and QASSO had diameter of 63.62 and 110.63 nm, and zeta potential of -68.89 and 48.25 mV at pH 7.0, respectively. Results indicated the GFP gene expression and cytoplasmic accumulation of GFP protein in the transformants after incubation under desirable conditions for 48 h and 1 week. The transformation efficiency was quantitatively assayed by flow cytometry, which yielded transformations of 58.87% and 48.34% for QASSO and 38.32% and a negligible percentage for Polyfect® after 48 h and 1 week incubation, respectively.
authors.affiliation: Research Center for Pharmaceutical Nanotechnology, Faculty of Pharmacy, Tabriz University of Medical Sciences, Tabriz, Iran; Student Research Committee, Faculty of Pharmacy, Tabriz University of Medical Sciences, Tabriz, Iran.
authors.forename: Fariba
authors.initials: F
authors.lastname: Akbari
authors.affiliation: Drug Applied Research Center, Faculty of Pharmacy, Tabriz University of Medical Sciences, Tabriz, Iran; Department of Pharmacognosy, Faculty of Pharmacy, Tabriz University of Medical Sciences, Tabriz, Iran. Electronic address: Yarikhosroushahia@tbzmed.ac.ir.
authors.forename: Ahmad
authors.initials: A
authors.lastname: Yari Khosroushahi
authors.affiliation: Iran Polymer and Petrochemical Institute (IPPI), P.O. Box 14965/115, Tehran, Iran. Electronic address: h.yeganeh@ippi.ac.ir.
authors.forename: Hamid
authors.initials: H
authors.lastname: Yeganeh
mesh_terms: D005434:Flow Cytometry; D015316:Genetic Therapy / Q000379:methods; D049452:Green Fluorescent Proteins / Q000737:chemistry* / Q000235:genetics; D058086:Microalgae / Q000737:chemistry*; D053758:Nanoparticles / Q000737:chemistry*; D010316:Particle Size; D000644:Quaternary Ammonium Compounds / Q000138:chemical synthesis* / Q000737:chemistry; D013024:Soybean Oil / Q000737:chemistry*; D014170:Transformation, Genetic
chemical_list: D000644:Quaternary Ammonium Compounds; C554142:epoxidized soybean oil; D049452:Green Fluorescent Proteins; D013024:Soybean Oil
keywords: Dunaliella salina; Green fluorescent protein; Nanolipoplex; Soybean oil