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Molecules, Vol. 23, Pages 1540: Synthesis and Comparative Evaluation of Novel Cationic Amphiphile C12-Man-Q as an Efficient DNA Delivery Agent In Vitro

Molecules, Vol. 23, Pages 1540: Synthesis and Comparative Evaluation of Novel Cationic Amphiphile C12-Man-Q as an Efficient DNA Delivery Agent In Vitro

Molecules doi: 10.3390/molecules23071540

Authors: Gunita Apsite Irena Timofejeva Aleksandra Vezane Brigita Vigante Martins Rucins Arkadij Sobolev Mara Plotniece Karlis Pajuste Tatjana Kozlovska Aiva Plotniece

New amphiphilic 1,4-DHP derivative C12-Man-Q with remoted cationic moieties at positions 2 and 6 was synthesised to study DNA delivery activity. The results were compared with data obtained for cationic 1,4-DHP derivative D19, which is known to be the most efficient one among the previously tested 1,4-DHP amphiphiles. We analysed the effects of C12-Man-Q concentration, complexation media, and complex/cell contact time on the gene delivery effectiveness and cell viability. Transmission electron microscopy data confirms that lipoplexes formed by the compound C12-Man-Q were quite uniform, vesicular-like structures with sizes of about 50 nm, and lipoplexes produced by compound D19 were of irregular shapes, varied in size in the range of 25–80 nm. Additionally, confocal microscopy results revealed that both amphiphiles effectively delivered green fluorescent protein expression plasmid into BHK-21 cells and produced a fluorescent signal with satisfactory efficiency, although compound C12-Man-Q was more cytotoxic to the BHK-21 cells with an increase of concentration. It can be concluded that optimal conditions for C12-Man-Q lipoplexes delivery in BHK-21 cells were the serum free media without 0.15 M NaCl, at an N/P ratio of 0.9. Compound D19 showed higher transfection efficiency to transfect BHK-21 and Cos-7 cell lines, when transfecting active proliferating cells. Although D19 was not able to transfect all studied cell lines we propose that it could be cell type specific. The compound C12-Man-Q showed modest delivery activity in all used cell lines, and higher activity was obtained in the case of H2-35 and B16 cells. The transfection efficiency in cell lines MCF-7, HeLa, and Huh-7 appears to be comparable to the reference compound D19 and minimal in the HepG2 cell line.

Authors:   Apsite, Gunita ; Timofejeva, Irena ; Vezane, Aleksandra ; Vigante, Brigita ; Rucins, Martins ; Sobolev, Arkadij ; Plotniece, Mara ; Pajuste, Karlis ; Kozlovska, Tatjana ; Plotniece, Aiva
Journal:   Molecules
Volume:   23
edition:   7
Year:   2018
Pages:   1540
DOI:   10.3390/molecules23071540
Publication date:   26-Jun-2018
Facts, background information, dossiers
  • cells
  • cell lines
  • transfection
  • molecules
  • DNA
  • concentration
  • green fluorescent protein
  • gene delivery
  • confocal microscopy
  • Complexation
  • cell viability
More about Molecular Diversity Preservation International
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