GlycofectionTM in the presence of anionic fusogenic peptides: a study of the parameters affecting the peptide‒mediated enhancement of the transfection efficiency

✍ Scribed by Antoine Kichler; Isabelle Freulon; Valérie Boutin; Roger Mayer; Michel Monsigny; Patrick Midoux

• Book ID: 101300199
• Publisher: John Wiley and Sons
• Year: 1999
• Tongue: English
• Weight: 317 KB
• Volume: 1
• Category: Article
• ISSN: 1099-498X
• DOI: 10.1002/(sici)1521-2254(199903/04)1:2<134::aid-jgm17>3.0.co;2-b

✦ Synopsis

Gene delivery mediated by polyplexes such as DNA complexed with polylysine conjugates is limited by the low ef®ciency of escape of DNA from the endosomes. One of the strategies which favors the transmembrane passage of polyplexes consists of adding anionic amphipathic peptides capable of destabilizing membranes in an acidic medium. Although less ef®cient than replication-defective adenoviruses, fusogenic peptides increase the expression of the reporter gene by a factor between 100 and 1000 depending on the cell line. However, the activity of a given peptide depends on the composition of the lipid bilayer. We were interested in developing a polyplex (glycoplex) formulation comprising a glycosylated polylysine, a fusogenic peptide and a plasmid which would be useful for ef®cient transfection (glycofection TM ) of a large panel of cells, even in the presence of serum. We synthesized several peptides and tested their ef®ciency in combination with different glycoplex formulations. We found that glycofection with a quaternary complex (called one pot formulation) made of lactosylated-polylysine, polylysine, DNA, and the dimeric peptide (E5-WYGG) 2 -KA was less cell-type dependent than other peptide-based formulations. In addition, its ef®ciency was not affected by the presence of serum (up to 20%).