Multidisciplinary Approach to the Transfection of Plasmid DNA by a Nonviral Nanocarrier Based on a Gemini-Bolaamphiphilic Hybrid Lipid

ACS Omega
María Martínez-NegroElena Junquera

Abstract

A multidisciplinary strategy, including both biochemical and biophysical studies, was proposed here to evaluate the potential of lipid nanoaggregates consisting of a mixture of a gemini-bolaamphiphilic lipid (C6C22C6) and the well-known helper lipid 1,2-dioleoyl-sn-glycero-3-phosphatidylethanolamine (DOPE) to transfect plasmid DNA into living cells in an efficient and safe way. For that purpose, several experimental techniques were employed, such as zeta potential (phase analysis light scattering methodology), agarose gel electrophoresis (pDNA compaction and pDNA protection assays), small-angle X-ray scattering, cryo-transmission electron microscopy, atomic force microscopy, fluorescence-assisted cell sorting, luminometry, and cytotoxicity assays. The results revealed that the cationic lipid and plasmid offer only 70 and 30% of their nominal positive () and negative charges (), respectively. Upon mixing with DOPE, they form lipoplexes that self-aggregate in typical multilamellar Lα lyotropic liquid-crystal nanostructures with sizes in the range of 100-200 nm and low polydispersities, very suitably fitted to remain in the bloodstream and cross the cell membrane. Interestingly, these nanoaggregates were able to compact, protect (...Continue Reading

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Citations

Oct 6, 2018·MedChemComm·M DamenM C Feiters
Feb 2, 2021·International Journal of Pharmaceutics·María L Santana-Armas, C Tros de Ilarduya
Feb 9, 2021·Frontiers in Chemistry·Jake R HughesPeter M Iovine

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Methods Mentioned

BETA
transfection
transfect
X-ray
atomic
electrophoresis
FACS
AFM
light scattering
Assay
fluorescence spectroscopy

Software Mentioned

DigitalMicrograph
BD CellQuest Pro
Quantity One
NanoScope Analysis

Related Concepts