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Higly fusogenic cationic liposomes transiently permeabilize the plasma membrane of HeLa cells

Abstract

Cationic liposomes can efficiently carry nucleic acids into mammalian cells. This property is tightly connected with their ability to fuse with negatively charged natural membranes (i.e. the plasma membrane and endosomal membrane). We used FRET to monitor and compare the efficiency of lipid mixing of two liposomal preparations — one of short-chained diC14-amidine and one of long-chained unsaturated DOTAP — with the plasma membrane of HeLa cells. The diC14-amidine liposomes displayed a much higher susceptibility to lipid mixing with the target membranes. They disrupted the membrane integrity of the HeLa cells, as detected using the propidium iodide permeabilization test. Morphological changes were transient and essentially did not affect the viability of the HeLa cells. The diC14-amidine liposomes were much more effective at either inducing lipid mixing or facilitating transfection.

Abbreviations

DiC14-amidine:

N-t-butyl-N’-tetradecylaminopropionamidine

DMPC:

dimyristoylphosphatidylcholine

DOTAP:

1,2-dioleoyl-3-trimethylammonium-propane

FRET:

fluorescence resonance energy transfer

MEMα:

minimum essential medium alpha

NBD-PE:

N-(7-nitrobenz-2-oxa-1,3-diazol-4-yl)-1,2-dihexadecanoyl-sn-3-phosphoetanolamine

PBS:

phosphate buffered saline

PC:

phosphatidylcholine

PMSF:

phenylmethylsulphonyl fluoride

PS:

phosphatidylserine

Rh-PE:

N-(lissamineTM rhodamine B sulfonyl)-1,2-dihexadecanoyl-sn-glycero-3-phospho-ethanolamine

SDS:

sodium dodecyl sulphate

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Stebelska, K., Wyrozumska, P., Gubernator, J. et al. Higly fusogenic cationic liposomes transiently permeabilize the plasma membrane of HeLa cells. Cell Mol Biol Lett 12, 39–50 (2007). https://doi.org/10.2478/s11658-006-0049-8

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  • DOI: https://doi.org/10.2478/s11658-006-0049-8

Key words

  • Cationic liposomes
  • Fusion
  • Transfection
  • Plasma membrane integrity