| Project/Area Number |
12672093
|
|---|
| Research Category |
Grant-in-Aid for Scientific Research (C)
|
| Allocation Type | Single-year Grants |
|---|
| Section | 一般 |
|---|
| Research Field |
Physical pharmacy
|
|---|
| Research Institution | Nagoya City University |
|---|
Principal Investigator |
HAZEMOTO Norio Nagoya City University, Associate Professor, 大学院・薬学研究科, 助教授 (40192273)
|
|---|
| Co-Investigator(Kenkyū-buntansha) |
YOTSUYANAGI Toshihisa Nagoya City University, Professor, 大学院・薬学研究科, 教授 (40080189)
四ツ柳 智久 名古屋市立大学, 薬学部, 教授 (06672150)
|
|---|
| Project Period (FY) |
2000 – 2001
|
| Project Status |
Completed (Fiscal Year 2001)
|
| Budget Amount *help |
¥3,400,000 (Direct Cost: ¥3,400,000)
Fiscal Year 2001: ¥900,000 (Direct Cost: ¥900,000)
Fiscal Year 2000: ¥2,500,000 (Direct Cost: ¥2,500,000)
|
|---|
| Keywords | gene delivery / DNA complex / oligo peptide / particle size / ternary complex / liposome / in vivo / stability / ペプチド合成 / 樹枝状ペプチド / プラスシドDNA / 遺伝子導入効率 / 核移行シグナル / 結合親和性 / 蛍光排除法 / 粒子径分布 / 原子間力顕微鏡 |
|---|
| 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.
|
