Polyethylenimine but not cationic lipids promotes transgene delivery to the nucleus in mammalian cells.
The beta-galactosidase reporter gene, either free or complexed with various cationic vectors, was microinjected into mammalian cells. Cationic lipids but not polyethylenimine or polylysine prevent transgene expression when complexes are injected in the nucleus. Polyethylenimine and to a lesser extent polylysine, but not cationic lipids, enhance transgene expression when complexes are injected into the cytoplasm. This latter effect was independent of the polymer vector/cDNA ionic charge ratio, suggesting that nucleic acid compaction rather than surface charge was critical for efficient nuclear trafficking. Cell division was not required for nuclear entry. Finally, comparative transfection and microinjection experiments with various cell lines confirm that barriers to gene transfer vary with cell type. We conclude that polymers but not cationic lipids promote gene delivery from the cytoplasm to the nucleus and that transgene expression in the nucleus is prevented by complexation with cationic lipids but not with cationic polymers.
Pollard
H
H
Laboratoire de Physiopathologie et de Pharmacologie Cellulaires et Moléculaires, INSERM CJF96-01, Hôpital Hotel-Dieu, Nantes, France.
Remy
J S
JS
Loussouarn
G
G
Demolombe
S
S
Behr
J P
JP
Escande
D
D
D000818:Animals; D019556:COS Cells; D002412:Cations; D002467:Cell Nucleus / Q000378:metabolism; D003593:Cytoplasm / Q000378:metabolism; D015870:Gene Expression; D006801:Humans; D050356:Lipid Metabolism; D008845:Microinjections; D011094:Polyethyleneimine / Q000378:metabolism*; D011107:Polylysine / Q000378:metabolism; D014162:Transfection / Q000379:methods*; D019076:Transgenes; D014407:Tumor Cells, Cultured
D002412:Cations; D011107:Polylysine; D011094:Polyethyleneimine