Role of cholesterol on the transfection barriers of cationic lipid/DNA complexes

Research output: Contribution to journalJournal articleResearchpeer-review

  • Daniela Pozzi
  • Francesco Cardarelli
  • Fabrizio Salomone
  • Cristina Marchini
  • Heinz Amenitsch
  • La Barbera, Giorgia
  • Giulio Caracciolo

Most lipid formulations need cholesterol for efficient transfection, but the precise motivation remains unclear. Here, we have investigated the effect of cholesterol on the transfection efficiency (TE) of cationic liposomes made of 1,2-dioleoyl-3-trimethylammonium-propane and dioleoylphosphocholine in Chinese hamster ovary cells. The transfection mechanisms of cholesterol-containing lipoplexes have been investigated by TE, synchrotron small angle X-ray scattering, and laser scanning confocal microscopy experiments. We prove that cholesterol-containing lipoplexes enter the cells using different endocytosis pathways. Formulations with high cholesterol content efficiently escape from endosomes and exhibit a lamellar-nonlamellar phase transition in mixture with biomembrane mimicking lipid formulations. This might explain both the DNA release ability and the high transfection efficiency. These studies highlight the enrichment in cholesterol as a decisive factor for transfection and will contribute to the rational design of lipid nanocarriers with superior TE.

Original languageEnglish
Article number073701
JournalApplied Physics Letters
Volume105
Issue number7
ISSN0003-6951
DOIs
Publication statusPublished - 18 Aug 2014

ID: 231311193