Uptake of exogenous DNA by rat liver: Effect of cationic lipids

R. Wattiaux, M. Jadot, F. Dubois, S. Misquith, S. Wattiaux-De Coninck

Research output: Contribution to journalArticlepeer-review

Abstract

We have investigated by using centrifugation methods, the uptake and the intracellular fate of S DNA by rat liver and the effect on these processes of N-(1-(2,3-dioleoxyloxy)propyl)-N,N,N,-trimethylammonium-methyl-sulfate(DOTAP, Boehringer, Mannheim, Germany), an artificial cationic lipid frequently used in transfection experiments. Labeled DNA molecules are quickly taken up by the liver but a progressive degradation takes place with time. Subcellular distribution of the radioactivity was established after differential and isopycnic centrifugation. Results indicate that S DNA enters liver cells by endocytosis and reaches lysosomes. The uptake of S DNA is not modified if the molecule is associated with DOTAP but marked differences are observed after internalization of the macromolecule. When DOTAP is used, radioactive products remain for a long time in low density organelles distinct from lysosomes indicating that the transfer of internalized DNA to these organelles is delayed by the cationic lipid. These results suggest that cationic lipids could favor transfection by preventing the delivery of DNA to lysosomes, allowing these molecules to be kept intact and available for transfer from endosomes to cytosol for a long time.
Original languageEnglish
Pages (from-to)81-87
Number of pages7
JournalBiochemical and Biophysical Research Communications
Volume213
Issue number1
DOIs
Publication statusPublished - 1 Jan 1995

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