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The role of endosome destabilizing activity in the gene transfer process mediated by cationic lipids
Author(s) -
Abdel El Ouahabi,
Marc Thiry,
Véronique Pector,
R. Fuks,
Jean Marie Ruysschaert,
Michel Vandenbranden
Publication year - 1997
Publication title -
febs letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.593
H-Index - 257
eISSN - 1873-3468
pISSN - 0014-5793
DOI - 10.1016/s0014-5793(97)00973-3
Subject(s) - endosome , cationic polymerization , transfection , endocytosis , chemistry , dna , plasmid , gene delivery , biophysics , cationic liposome , biochemistry , biology , intracellular , gene , organic chemistry , cell
We used a 32 P‐labeled pCMV‐CAT plasmid DNA to estimate the DNA uptake efficiency and unlabeled pCMV‐CAT plasmid DNA to quantify the CAT activity after transfection of COS cells using each of the three following cationic compounds: [1] vectamidine (3‐tetradecylamino‐ N ‐tert‐butyl‐ N ′‐tetradeccyl‐propionamidine, and previously described as diC14‐amidine [1]), [2] lipofectin (a 1:1 mixture of N ‐(1‐2,3‐dioleyloxypropyl) N,N,N ‐triethylammonium (DOTMA) and dioleylphosphatid‐ylethanolamine (DOPE)), and [3] DMRIE‐C (a 1:1 mixture of N ‐[1‐(2,3‐dimyristyloxy)propyl]‐ N,N ‐dimethyl‐ N ‐(2‐hydroxyethyl) ammonium bromide (DMRIE) and cholesterol). Surprisingly, a high CAT activity was observed with vectamidine although the DNA uptake efficiency was lower as compared to lipofectin and DMRIE‐C. Transmission electron microscopy (TEM) revealed endocytosis as the major pathway of DNA‐cationic lipid complex entry into COS cells for the three cationic lipids. However, the endosomal membrane in contact with complexes containing vectamidine or DMRIE‐C often exhibited a disrupted morphology. This disruption of endosomes was much less frequently observed with the DNA‐lipofectin complexes. This comparison of the three compounds demonstrate that efficient transfection mediated by cationic lipids is not only correlated to their percentage of uptake but also to their ability to destabilize and escape from endosomes.