title: (LYS)(16)-based reducible polycations provide stable polyplexes with anionic fusogenic peptides and efficient gene delivery to post mitotic cells.
abstract: Extracellular stability, endocytic escape, intracellular DNA release and nuclear translocation of DNA are all critical properties of non-viral vector/DNA particles. We have evaluated a (Lys)(16)-based linear, reducible polycation (RPC) in combination with an acid-dependent, anionic fusogenic peptide for gene delivery to dividing and post-mitotic cells. The RPC was formed from Cys(Lys)(16)Cys monomers. Molecular weight was 24,000 Da, corresponding to an average of 10.5 peptide monomers per RPC. Non-reducible polylysine (PLL) (27,000 Da) and monomeric (Lys)(16) peptide were evaluated for comparison. (Lys)(16)/DNA particles were disrupted at fusogenic peptide concentrations well below those used for gene delivery. By contrast, RPC/DNA an PLL/DNA particles were stable in the presence of high concentrations of the anionic peptide. Addition of 10% serum virtually abolished the transfection ability of (Lys)(16)/DNA/fusogenic peptide particles, but had little effect on RPC/DNA/fusogenic peptide particles. RPC/DNA/fusogenic peptide particles were highly effective for gene delivery to both cell lines and post-mitotic corneal endothelium. PLL/DNA/fusogenic peptide particles were moderately effective on cell lines, but gave no gene delivery with corneal endothelial cells. We conclude that (Lys)(16)-based RPC/DNA/fusogenic peptide particles provide a gene delivery system which is potentially stable in the extracellular environment and, on reductive depolymerisation, can release DNA plasmids for nuclear translocation.
authors.affiliation: Department of Clinical Sciences King's College London School of Medicine The Rayne Institute 123 Coldharbour Lane, London SE5 9NU, UK.
authors.forename: Alan L
authors.initials: AL
authors.lastname: Parker
authors.forename: Lorna
authors.initials: L
authors.lastname: Eckley
authors.forename: Surjeet
authors.initials: S
authors.lastname: Singh
authors.forename: Jon A
authors.initials: JA
authors.lastname: Preece
authors.forename: Louise
authors.initials: L
authors.lastname: Collins
authors.forename: John W
authors.initials: JW
authors.lastname: Fabre
mesh_terms: D000256:Adenoviridae; D000818:Animals; D002460:Cell Line; D003315:Cornea / Q000166:cytology; D018014:Gene Transfer Techniques; D005822:Genetic Vectors; D006367:HeLa Cells; D006801:Humans; D008297:Male; D008561:Membrane Fusion; D053758:Nanoparticles; D010455:Peptides / Q000008:administration & dosage*; D011073:Polyamines / Q000008:administration & dosage*; D000071228:Polyelectrolytes; D011107:Polylysine / Q000008:administration & dosage*; D011817:Rabbits; D016192:Resting Phase, Cell Cycle
chemical_list: D010455:Peptides; D011073:Polyamines; D000071228:Polyelectrolytes; D011107:Polylysine