Novel Cationic Phosphonolipids Agents for in Vitro Gene Transfer to Epithelial Cell Lines

Abstract

The use of cationic liposomes is one of the main approaches currently investigated to introduce into a cell a gene with therapeutic properties. This study presents in vitro results obtained with a new family of gene transfer agents, the phosphonolipids. We have synthesized 37 members of this family and optimized the conditions of in vitro gene transfer targeted at a lung epithelial cell line (CFT1 cells) by using a reporter gene (β-galactosidase). Two quantitative tests, a CPRG¹ (Chlorophenol red galactopyranoside) test and a Flow cytometric assay (FACS-Gal¹ assay), have been used to determine the percentage of transfected cells. The cationic phosphonolipids were tested alone or formulated with 50% DOPE¹ (Dioleoylphosphatidylethanolamine) (w:w). The results obtained with the CPRG test led us to select 7 compounds that were more efficient than the commercialized lipids Lipofectin, Lipofectamine and Transfectam. The transfer kinetics of the transgene were studied and showed that more than 20 % of cells were still positive at day 7 after transfection.