Gene delivery by negatively charged ternary complexes of DNA, cationic liposomes and transferrin or fusigenic peptides

Abstract

Potential problems with the use of viral vectors for gene also enhanced by association with transferrin, GALA or the therapy necessitate the development of efficient nonviral influenza hemagglutinin N terminal peptide HA-2, but to a vectors. The association of transferrin, or the pH-sensitive smaller extent compared with the negatively charged compeptide GALA, with cationic liposomes composed of 1,2-plexes. The enhancement of gene delivery by transferrin dioleoyl-3-(trimethylammonium) propane and its equimolar or GALA was not affected significantly by the presence of mixture with dioleoylphosphatidylethanolamine, under conserum and did not cause significant cytotoxicity. Our ditions where the liposome/DNA complex is negatively results indicate that negatively charged ternary complexes charged, drastically increased luciferase expression from of cationic liposomes, DNA and transferrin, or fusigenic pCMVluc. The percentage of cells transfected, measured peptides, can facilitate efficient transfection of cultured by ␤-galactosidase expression, was also increased by cells, and that they may alleviate the drawbacks of the use about 10-fold. The zeta potential of the ternary complexes of highly positively charged complexes for gene delivery was lower than that of the liposome/DNA complexes.in vivo. Transfection activity of positively charged complexes was

Improving lipoplex-mediated gene transfer into C6 glioma cells and primary neurons

“…Coating of lipoplexes with Tf has been demonstrated to promote both the cellular uptake of lipoplexes (da Cruz et al, 2001) and to trigger cytoplasmic delivery of the carried DNA through destabilization of the endosomal membrane under acidic conditions (da Cruz et al, 2001;Simoes et al, 1998). Our results clearly indicate that such a strategy resulted in a significant enhancement of transfection in both neuronal primary cultures and cell line, this effect being dependent on the type of cells, net charge, and composition of the complexes.…”

“…Targeting DNA vectors to cellular Tf receptors has been tried successfully before by linking the ligand to polycation conjugates (Liu et al, 1995;Zhu et al, 1993), through association of the protein with cationic liposomes and by covalent coupling of anti-Tf receptor monoclonal antibodies to the distal ends of PEG on sterically stabilized liposomes (Shi et al, 2001) or on nanoparticles (Olivier et al, 2002). In this regard, we and others have demonstrated that the association of human Tf to cationic liposome/DNA complexes enhances transfection in a large variety of cells, including dividing and nondividing cells (Cheng, 1996;Simoes et al, 1998Simoes et al, , 1999b.…”

“…It has been demonstrated that Tf is highly effective in promoting lipoplex-mediated transfection of various cells lines and primary cell cultures Simoes et al, 1998Simoes et al, , 1999aWagner et al, 1994). Therefore, in this work, we evaluated whether this strategy would result in an enhancement of transfection of neuronal cells.…”

“…Aiming at enhancing transfection activity mediated by cationic liposomes, different strategies have been tried, including the association of targeting ligands (Cheng, 1996;Simoes et al, 1998Simoes et al, , 1999b, fusogenic proteins (Simoes et al, 2000), and fusogenic (Kichler et al, 1997;Plank et al, 1994) and NLS (Chan and Jans, 1999;Zanta et al, 1999) peptides. Targeting DNA vectors to cellular Tf receptors has been tried successfully before by linking the ligand to polycation conjugates (Liu et al, 1995;Zhu et al, 1993), through association of the protein with cationic liposomes and by covalent coupling of anti-Tf receptor monoclonal antibodies to the distal ends of PEG on sterically stabilized liposomes (Shi et al, 2001) or on nanoparticles (Olivier et al, 2002).…”

Single-chain antibody-mediated gene delivery into ErbB2-positive human breast cancer cells

“…The size and zeta potential of the particles, although not studied here, can be subsequently measured and the individual components of the particles adjusted to optimize in vivo delivery. 35 The DPSL complex demonstrates advantages over ScFv -P -S including ease in large -scale preparation in addition to higher transfection efficiency while substantially retaining target selectivity.…”