RT Journal Article
T1 Multidisciplinary approach to the transfection of plasmid DNA by a nonviral nanocarrier based on a Gemini-Bolaamphiphilic hybrid lipid
A1 Martínez Negro, María
A1 Guerrero Martínez, Andrés
A1 García Río, Luis
A1 Domènech, Òscar
A1 Aicart, Emilio
A1 Tros de Ilarduya, Conchita
A1 Junquera, Elena
K1 Vesicles
K1 Lipids
K1 Cytosine
K1 Genetics
K1 Chemical structure
AB A multidisciplinary strategy, including bothbiochemical and biophysical studies, was proposed here toevaluate the potential of lipid nanoaggregates consisting of amixture of a gemini−bolaamphiphilic lipid (C6C22C6) and thewell-known helper lipid 1,2-dioleoyl-sn-glycero-3-phosphatidy-lethanolamine (DOPE) to transfect plasmid DNA into livingcells in an efficient and safe way. For that purpose, severalexperimental techniques were employed, such as zeta potential(phase analysis light scattering methodology), agarose gelelectrophoresis (pDNA compaction and pDNA protectionassays), small-angle X-ray scattering, cryo-transmission electronmicroscopy, atomic force microscopy,fluorescence-assisted cellsorting, luminometry, and cytotoxicity assays. The resultsrevealed that the cationic lipid and plasmid offer only 70 and30% of their nominal positive (=++q2.0nom,C C  C6226) and negative charges (=−−q2/bpnom,pDNA), respectively. Upon mixing withDOPE, they form lipoplexes that self-aggregate in typical multilamellar Lαlyotropic liquid-crystal nanostructures with sizes in therange of 100−200 nm and low polydispersities, very suitablyfitted to remain in the bloodstream and cross the cell membrane.Interestingly, these nanoaggregates were able to compact, protect (from the degrading effect of DNase I), and transfect two DNAplasmids (pEGFP-C3, encoding the greenfluorescent protein, and pCMV-Luc, encoding luciferase) into COS-7 cells, with anefficiency equal or even superior to that of the universal control Lipo2000*, as long as the effective +/−charge ratio wasmaintained higher than 1 but reasonably close to electroneutrality. Moreover, this transfection process was not cytotoxic becausethe viability of COS-7 cells remained at high levels, greater than 80%. All of these features make the C6C22C6/DOPE nanosysteman optimal nonviral gene nanocarrier in vitro and a potentially interesting candidate for future in vivo experiments
PB American Chemical Society
SN 2470-1343
YR 2018
FD 2018
LK http://hdl.handle.net/10347/23202
UL http://hdl.handle.net/10347/23202
LA eng
NO ACS Omega 2018, 3, 1, 208–217
NO Financial support from the Ministerio de Economia y Competitividad of Spain (projects CTQ2012-30821, CTQ2015-65972-R, CTQ2015-64425-C2-2-R, and CTQ2014-55208-P), Madrid Regional Government (S2013/MIT-2807), Xunta de Galicia (GR 2007/085; IN607C 2016/03 and Centro Singular de Investigación de Galicia accreditation 2016–2019, ED431G/09), the European Regional Development Fund (ERDF), and Universidad Complutense de Madrid, Spain (project UCMA05-33-010), is gratefully acknowledged
DS Minerva
RD 25 abr 2026