Effect of oligopeptides on gene expression: comparison of DNA/peptide and DNA/peptide/liposome complexes.
Plasmid DNA is known to form complexes with a variety of cationic peptides and lipids, which have been explored as possible carriers for DNA transfection in mammalian cells. We synthesized oligopeptides consisting of nine amino acid residues including lysine (K), tryptophan (W), and cysteine (C), and also their symmetrical dimmers with a disulfide bond as possible carriers. The pDNA(pGL3)/oligopeptide complexes generally showed poor transfection efficiencies but little cytotoxicity for HeLa S3. The ternary system of pDNA/oligopeptide/liposome containing cationic liposomes formulated from the cholesterol derivative (DMB-Chol) and dioleoylphosphatidylethanolamine (DOPE) showed 10(4)-10(5)-fold greater effective gene expression (10(8)-10(9) level, RLU/min/mg protein) than those of the corresponding pDNA/oligopeptide complexes. In the presence of 10% serum, the ternary complexes were maintained at 10(7) levels. The ethidium bromide exclusion studies showed the ternary complexes have much greater affinity to pDNA than the corresponding pDNA/oligopeptide complexes. Plasmid sensitivity against DNase I degradation showed that the ternary complexes were well protected from the digestion. Synthetic oligopeptides are active as potential enhancers for DOPE-containing cationic liposome-mediated transfection. These findings have implications for successful in vivo transfection.
Tokunaga
Masaki
M
Department of Pharmaceutics, Graduate School of Pharmaceutical Sciences, Nagoya City University, Tanabe-dori, Mizuho-ku, Nagoya 467-8603, Japan.
Hazemoto
Norio
N
Yotsuyanagi
Toshihisa
T
D000440:Butanols / Q000737:chemistry; D002784:Cholesterol / Q000031:analogs & derivatives; D004247:DNA / Q000008:administration & dosage* / Q000737:chemistry; D003850:Deoxyribonuclease I / Q000737:chemistry; D004587:Electrophoresis, Agar Gel; D004996:Ethidium / Q000737:chemistry; D015870:Gene Expression / Q000187:drug effects*; D017930:Genes, Reporter; D006367:HeLa Cells; D006801:Humans; D008081:Liposomes; D008156:Luciferases / Q000235:genetics / Q000378:metabolism; D009842:Oligopeptides / Q000737:chemistry / Q000494:pharmacology*; D010316:Particle Size; D010714:Phosphatidylethanolamines; D010957:Plasmids; D014162:Transfection
C484936:4-dimetylamino-1-butanol; D000440:Butanols; D008081:Liposomes; D009842:Oligopeptides; D010714:Phosphatidylethanolamines; C094877:dioleoyl phosphatidylethanolamine; D004247:DNA; D002784:Cholesterol; D008156:Luciferases; D003850:Deoxyribonuclease I; D004996:Ethidium