Artículo
A Non-Viral Plasmid DNA Delivery System Consisting on a Lysine-Derived Cationic Lipid Mixed with a Fusogenic Lipid
Autor/es | Martínez Negro, María
Sánchez Arribas, Natalia Guerrero Martínez, Andrés Moyá Morán, María Luisa Tros de Llarduya, Conchita Mendicuti, Francisco Aicart, Emilio Junquera, Elena |
Departamento | Universidad de Sevilla. Departamento de Química Física |
Fecha de publicación | 2019 |
Fecha de depósito | 2019-12-02 |
Publicado en |
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Resumen | The 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 ... The 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* |
Identificador del proyecto | contract numbers CTQ2015-65972-R, CTQ2015-64425-C2-1-R, CTQ2015-64425-C2-2-R, CTQ2016-80600-P and RTI2018-095844-B-I00)
project number UCMA05-33-010 project number CCGP2017-EXP/027 |
Cita | Martínez Negro, M., Sánchez Arribas, N., Guerrero Martínez, A., Moyá Morán, M.L., Tros de Llarduya, C., Mendicuti, F.,...,Junquera, E. (2019). A Non-Viral Plasmid DNA Delivery System Consisting on a Lysine-Derived Cationic Lipid Mixed with a Fusogenic Lipid. Pharmaceutics, 11 (12), 1-16. |
Ficheros | Tamaño | Formato | Ver | Descripción |
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pubpharmaceutics-11-00632.pdf | 3.120Mb | [PDF] | Ver/ | |