19 research outputs found
A gemini cationic lipid with histidine residues as a novel lipid-based gene nanocarrier: a biophysical and biochemical study
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-glycero3-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
A Non-Viral Plasmid DNA Delivery System Consisting on a Lysine-Derived Cationic Lipid Mixed with a Fusogenic Lipid
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*
Trehalose-based siamese twin amphiphiles with tunable self-assembling, DNA nanocomplexing and gene delivery properties
An original family of multivalent vectors encompassing gemini and facial amphiphilicity, namely cationic Siamese twin surfactants, has been prepared fromthe disaccharide trehalose; molecular engineering lets us modulate the self-assembling properties and the topology of the nanocomplexes with plasmid DNA for efficient gene delivery in vitro and in vivo