TY - JOUR
T1 - Uptake of exogenous DNA by rat liver
T2 - Effect of cationic lipids
AU - Wattiaux, R.
AU - Jadot, M.
AU - Dubois, F.
AU - Misquith, S.
AU - Wattiaux-De Coninck, S.
N1 - Medline is the source for the MeSH terms of this document.
PY - 1995/1/1
Y1 - 1995/1/1
N2 - 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.
AB - 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.
UR - http://www.scopus.com/inward/record.url?scp=0029075867&partnerID=8YFLogxK
U2 - 10.1006/bbrc.1995.2101
DO - 10.1006/bbrc.1995.2101
M3 - Article
AN - SCOPUS:0029075867
SN - 0006-291X
VL - 213
SP - 81
EP - 87
JO - Biochemical and Biophysical Research Communications
JF - Biochemical and Biophysical Research Communications
IS - 1
ER -