2001 Fiscal Year Final Research Report Summary
Gene delivery mediated by synthetic oligopeptide
| Project/Area Number |
12672093
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| Research Category |
Grant-in-Aid for Scientific Research (C)
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
Physical pharmacy
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| Research Institution | Nagoya City University |
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Principal Investigator |
HAZEMOTO Norio Nagoya City University, Associate Professor, 大学院・薬学研究科, 助教授 (40192273)
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| Co-Investigator(Kenkyū-buntansha) |
YOTSUYANAGI Toshihisa Nagoya City University, Professor, 大学院・薬学研究科, 教授 (40080189)
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| Project Period (FY) |
2000 – 2001
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| Keywords | gene delivery / DNA complex / oligo peptide / particle size / ternary complex / liposome / in vivo / stability |
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| Research Abstract |
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 DNA/oligopeptide/liposome containing cationic liposome formulated from the cholesterol derivative (DMB-Chol) and dioleylphosphatidylethanolamine (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 level. The ethidium bromide exclusion studies showed the ternary complexes have much greater affinity to pDNA than the corresponding pDNA/oligopeptide complexes. Plasmid sensitivity against DnaseI 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.
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