Top of pageAbstract Over the last decade, several barriers have been overcome in the field of DNA technology, leading to its consideration for use in gene therapy as a possible treatment for many diseases. However, the major problem is still the same: that is, getting efficient and safe carriers to transfer the gene to specific cells. Even though the virus is being used in most clinical trials because of high transfection efficiency, recent results highlight many concerns about its use, especially immunogenicity and random integration. In parallel, cationic reagents have progressed spectacularly from their first transfections. Nevertheless, for synthetic reagents, one of the limitations is still their relatively low transfection efficiency. In order to increase their efficiency, one approach could be the cellular targeting. Previous research into gene transfer delivery via sugar receptors appears interesting; however glycotargeting is still in its early stages. Indeed, current knowledge of the functions and expression of lectine receptors is incomplete and represent a limiting factor to their potential use. Even if this strategy has been already succeed, effective receptors mediated endocytosis phenomenon depends on several conditions (Expression of the receptor, affinity for the ligand, size of the lipoplex, immune response …). These difficulties led to the use of smaller ligands, such as synthetic oligosaccharides, and it has been reported some efficient transfection results using plasmid DNA complexed with Poly-L-Lysine bearing either lactosyl or mannosyl residues. However, considering polymers, mainly adapted from Poly-L-lysine or PEI, a high toxicity was observed both in vitro and in vivo. For CF treatment, gene transfer should specifically target airway surface ciliated cells and gland serous cells in order to induce the expression of a normal CFTR protein at the apical membrane of CF cells. For targeted gene delivery, we extended our studies by conjugating a glycosylated residue (lactose or mannose) to quaternary ammonium lipids possessing branched and/or linear chains or neutral lipids composed of a couple of hydrophobic chains. Several combinations of these glycosylated amphiphiles were used in the lipoplexes preparations, with or without the presence of a co-lipid as DOPE or cholesterol. Then, these formulations were tested in vitro on two human cell lines (Hela and 16HBE14o(-)). We report here that one of those formulations (β-D-Galactopyranosyl-(1,4)-N-β-D-glucopyranosyl-(N,N-dimethyl-N-hexadecyl-3-ammoniumyl)propylhexadecanamide bromide/DOPE) is more efficient than DOTAP/DOPE. We determined this transfection is partially due to an endocytotic phenomenon mediated by targeting specific receptors directed towards specific carbohydrate elements. This was shown on 16HBE14o(-) cells where we observed a forty three and a sixty nine percent decrease in transfection when we blocked these receptors by the addition of free lactose and mannose respectively. These results highlight that transfection is mediated by specific receptors, especially the sugar receptors, and using cationic glycolipids, instead of polymers agents, is encouraging.