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DNA-DOPE-gemini surfactants complexes at low surface charge density: from structure to transfection efficiency
Lukáš Hubčík1, Dominika Galliková, Petra Pullmannová
1Department of Physical Chemistry of Drugs, Faculty of Pharmacy, Comenius University in Bratislava, Odbojárov 10, 832 32 Bratislava, Slovakia. hubcik@fpharm.uniba.sk.
General Physiology and Biophysics
|February 10, 2018
Summary
Gemini surfactants with dioleoylphosphatidylethanolamine (DOPE) and DNA form condensed complexes. The C12GS12/DOPE/DNA complex showed the highest transfection efficiency in HEK 293 cells, with good cell viability.
Area of Science:
- Biomaterials Science
- Nanotechnology
- Molecular Biology
Background:
- Gemini surfactants are amphiphilic molecules with potential applications in gene delivery.
- Understanding the structure-activity relationship of gemini surfactant-DNA complexes is crucial for optimizing transfection efficiency.
Purpose of the Study:
- To investigate the DNA condensation, structure, and transfection efficiency of complexes formed by gemini surfactants (CnGS12), dioleoylphosphatidylethanolamine (DOPE), and DNA.
- To evaluate the effect of the alkane spacer length in gemini surfactants on these properties.
Main Methods:
- Small angle X-ray diffraction (SAXD) to study DNA condensation and structure.
- Fluorescence and UV-VIS spectroscopy to determine the percentage of complexed DNA.
- In vitro transfection assays on HEK 293 cell line to assess transfection efficiency and cell viability.
Main Results:
- Gemini surfactants, DOPE, and DNA formed condensed lamellar phases, with DNA-DNA distance dependent on the charge ratio.
- High ionic strength reduced DNA-surfactant interactions; 45-60% of DNA was complexed.
- Transfection efficiency was spacer length-dependent, with C12GS12/DOPE/DNA complexes showing the highest efficiency (~18% GFP-expressing cells) and good cell viability (~89%).
Conclusions:
- Gemini surfactants, in combination with DOPE, can effectively condense DNA for gene delivery applications.
- The spacer length of gemini surfactants significantly influences transfection efficiency and cell viability.
- C12GS12/DOPE/DNA complexes represent a promising non-viral vector for gene transfection.
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