RT Journal Article
T1 A Non-Viral Plasmid DNA Delivery System Consisting on a Lysine-Derived Cationic Lipid Mixed with a Fusogenic Lipid
A1 Martínez Negro, María
A1 Sánchez Arribas, Natalia
A1 Guerrero Martínez, Andrés
A1 Moyá, María Luisa
A1 Tros de Ilarduya, Conchita
A1 Mendicuti, Francisco
A1 Aicart Sospedra, Emilio
A1 Junquera González, Elena
AB 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*.
PB MDPI
SN 1999-4923
YR 2019
FD 2019-11-27
LK https://hdl.handle.net/20.500.14352/8281
UL https://hdl.handle.net/20.500.14352/8281
LA eng
NO Ministerio de Economía y Competitividad (MINECO)
NO Universidad Complutense de Madrid
NO Universidad de Alcalá
DS Docta Complutense
RD 27 abr 2024