Martínez Negro, MaríaSánchez Arribas, NataliaGuerrero Martínez, AndrésMoyá, María LuisaTros de Ilarduya, ConchitaMendicuti, FranciscoAicart Sospedra, EmilioJunquera González, Elena2023-06-172023-06-172019-11-271999-492310.3390/pharmaceutics11120632https://hdl.handle.net/20.500.14352/8281The insertion of biocompatible amino acid moieties in non-viral gene nanocarriers is an attractive approach that has been recently gaining interest. In this work, a cationic lipid, consisting of a lysine-derived moiety linked to a C12 chain (LYCl) was combined with a common fusogenic helper lipid (DOPE) and evaluated as a potential vehicle to transfect two plasmid DNAs (encoding green fluorescent protein GFP and luciferase) into COS-7 cells. A multidisciplinary approach has been followed: (i) biophysical characterization based on zeta potential, gel electrophoresis, small-angle X-ray scattering (SAXS), and cryo-transmission electronic microscopy (cryo-TEM); (ii) biological studies by fluorescence assisted cell sorting (FACS), luminometry, and cytotoxicity experiments; and (iii) a computational study of the formation of lipid bilayers and their subsequent stabilization with DNA. The results indicate that LYCl/DOPE nanocarriers are capable of compacting the pDNAs and protecting them efficiently against DNase I degradation, by forming Lα lyotropic liquid crystal phases, with an average size of ~200 nm and low polydispersity that facilitate the cellular uptake process. The computational results confirmed that the LYCl/DOPE lipid bilayers are stable and also capable of stabilizing DNA fragments via lipoplex formation, with dimensions consistent with experimental values. The optimum formulations (found at 20% of LYCl content) were able to complete the transfection process efficiently and with high cell viabilities, even improving the outcomes of the positive control Lipo2000*.engAtribución 3.0 Españajournal articlehttps://doi.org/10.3390/pharmaceutics11120632https://www.mdpi.com/1999-4923/11/12/632open accesslysine-derived cationic lipidplasmid DNAlipoplextransfectionprotectioncompactiongene deliverymultilamellar aggregatesmolecular dynamicsQuímica física (Física)Química física (Química)2210 Química Física