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Glycofection TM in the presence of anionic fusogenic peptides: a study of the parameters affecting the peptide‒mediated enhancement of the transfection efficiency
Author(s) -
Kichler Antoine,
Freulon Isabelle,
Boutin Valérie,
Mayer Roger,
Monsigny Michel,
Midoux Patrick
Publication year - 1999
Publication title -
the journal of gene medicine
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.689
H-Index - 91
eISSN - 1521-2254
pISSN - 1099-498X
DOI - 10.1002/(sici)1521-2254(199903/04)1:2<134::aid-jgm17>3.0.co;2-b
Subject(s) - polylysine , peptide , transfection , gene delivery , chemistry , biophysics , amphiphile , reporter gene , endosome , dna , biochemistry , cell , biology , gene , gene expression , organic chemistry , copolymer , polymer
Gene delivery mediated by polyplexes such as DNA complexed with polylysine conjugates is limited by the low efficiency 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 efficient 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 efficient transfection (glycofection TM ) of a large panel of cells, even in the presence of serum. We synthesized several peptides and tested their efficiency 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 efficiency was not affected by the presence of serum (up to 20%). Copyright © 1999 John Wiley & Sons, Ltd.