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A gemini cationic lipid with histidine residues as a novel lipid-based gene nanocarrier: A biophysical and biochemical study

AuthorsMartínez-Negro, María; Blanco-Fernández, Laura; Tentori, Paolo M.; Pérez Muñoz, Lourdes; Pinazo Gassol, Aurora; Tros de Ilarduya, Conchita; Aicart, Emilio; Junquera, Elena
KeywordsLipid-based gene nanocarrier
Gemini cationic lipid with histidine residues
Gene delivery
Plasmid DNAs
Cell viabilit
Biophysical characterization
Issue Date16-Dec-2018
PublisherMultidisciplinary Digital Publishing Institute
CitationNanomaterials 8(12): 1061 (2018)
AbstractThis 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.
Publisher version (URL)https://doi.org/10.3390/nano8121061
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