A Gemini Cationic Lipid with Histidine Residues as a Novel Lipid-Based Gene Nanocarrier: A Biophysical and Biochemical Study

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Martínez Negro, María and Blanco-Fernández, Laura and Tentori, Paolo M. and Pérez, Lourdes and Pinazo, Aurora and Tros de Ilarduya, Conchita and Aicart Sospedra, Emilio and Junquera González, Elena (2018) A Gemini Cationic Lipid with Histidine Residues as a Novel Lipid-Based Gene Nanocarrier: A Biophysical and Biochemical Study. Nanomaterials, 8 (12). p. 1061. ISSN 2079-4991

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Official URL: https://doi.org/10.3390/nano8121061




Abstract

This work reports the synthesis of a novel gemini cationic lipid that incorporates two histidine-type head groups (C3(C16His)2). Mixed with a helper lipid 1,2-dioleoyl-sn-glycero-3-phosphatidyl ethanol amine (DOPE), it was used to transfect three different types of plasmid DNA: one encoding the green fluorescence protein (pEGFP-C3), one encoding a luciferase (pCMV-Luc), and a therapeutic anti-tumoral agent encoding interleukin-12 (pCMV-IL12). Complementary biophysical experiments (zeta potential, gel electrophoresis, small-angle X-ray scattering (SAXS), and fluorescence anisotropy) and biological studies (FACS, luminometry, and cytotoxicity) of these C3(C16His)2/DOPE-pDNA lipoplexes provided vast insight into their outcomes as gene carriers. They were found to efficiently compact and protect pDNA against DNase I degradation by forming nanoaggregates of 120–290 nm in size, which were further characterized as very fluidic lamellar structures based in a sandwich-type phase, with alternating layers of mixed lipids and an aqueous monolayer where the pDNA and counterions are located. The optimum formulations of these nanoaggregates were able to transfect the pDNAs into COS-7 and HeLa cells with high cell viability, comparable or superior to that of the standard Lipo2000*. The vast amount of information collected from the in vitro studies points to this histidine-based lipid nanocarrier as a potentially interesting candidate for future in vivo studies investigating specific gene therapies.


Item Type:Article
Uncontrolled Keywords:lipid-based gene nanocarrier; gemini cationic lipid with histidine residues; gene delivery; plasmid DNAs; transfection; cell viability; biophysical characterization
Subjects:Sciences > Physics > Chemistry, Physical and theoretical
Sciences > Chemistry > Chemistry, Physical and theoretical
ID Code:66760
Deposited On:19 Jul 2021 14:33
Last Modified:20 Jul 2021 07:18

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