The influence of endosome-disruptive peptides on gene transfer using synthetic virus-like gene transfer systems.
The process by which viruses destabilize endosomal membranes in an acidification-dependent manner has been mimicked with synthetic peptides that are able to disrupt liposomes, erythrocytes, or endosomes of cultured cells. Peptides containing the 20 amino-terminal amino acid sequence of influenza virus hemagglutinin as well as acidic derivatives showed erythrocyte lysis activity only when peptides were elongated by an amphipathic helix or by carboxyl-terminal dimerization. Interestingly, peptides consisting of the 23 amino-terminal amino acids of influenza virus hemagglutinin were also active in erythrocyte lysis. When peptides were incorporated into DNA complexes that utilize a receptor-mediated endocytosis pathway for uptake into cultured cells, either by ionic interaction with positively charged polylysine-DNA complexes or by a streptavidin-biotin bridge, a strong correlation between pH-specific erythrocyte disruption activity and gene transfer was observed. A high-level expression of luciferase or interleukin-2 was obtained with optimized gene transfer complexes in human melanoma cells and several cell lines.
Plank
C
C
Research Institute of Molecular Pathology, Vienna, Austria.
Oberhauser
B
B
Mechtler
K
K
Koch
C
C
Wagner
E
E
D000595:Amino Acid Sequence; D002478:Cells, Cultured; D004705:Endocytosis; D018014:Gene Transfer Techniques; D005814:Genes, Viral; D019267:Hemagglutinin Glycoproteins, Influenza Virus; D006389:Hemagglutinins, Viral / Q000737:chemistry; D006801:Humans; D008969:Molecular Sequence Data; D015388:Organelles / Q000378:metabolism*; D010455:Peptides / Q000138:chemical synthesis / Q000494:pharmacology*; D014162:Transfection; D014759:Viral Envelope Proteins / Q000737:chemistry
D019267:Hemagglutinin Glycoproteins, Influenza Virus; D006389:Hemagglutinins, Viral; D010455:Peptides; D014759:Viral Envelope Proteins