Gene delivery mediated by synthetic oligopeptide

合成寡肽介导的基因传递

• 批准号: 12672093
• 负责人: HAZEMOTO Norio
• 金额: $ 2.18万
• 依托单位: Nagoya City University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 2000
• 资助国家: 日本
• 起止时间: 2000 至 2001
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-12672093/
• 关键词: 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

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.

质粒DNA已知与多种阳离子肽和脂质形成复合物，已被探索作为哺乳动物细胞中DNA转染的可能载体。我们合成了由9个氨基酸残基组成的寡肽，包括赖氨酸(K)、色氨酸(W)和半胱氨酸(C)，以及它们对称的二硫键二聚体作为可能的载体。pDNA (pGL3)/寡肽复合物对HeLa S3的转染效率一般较差，但细胞毒性较小。由胆固醇衍生物（DMB-Chol）和二油基磷脂酰乙醇胺（DOPE）组成的含有阳离子脂质体的DNA/寡肽/脂质体三元体系的有效基因表达量（10^8-10^9水平，RLU/min/mg蛋白）比相应的pDNA/寡肽复合物高10^4-10^5倍。在10%血清存在的情况下，三元复合物维持在10^7水平。溴化乙啶的排除研究表明，三元配合物对pDNA的亲和力远高于相应的pDNA/寡肽配合物。质粒对dna i降解的敏感性表明，三元配合物受到很好的保护，不受消化的影响。人工合成的寡肽在阳离子脂质体介导的dope转染中具有潜在的增强作用。这些发现对成功的体内转染具有启示意义。

项目成果

Norio Hazemoto et al.: "Gene delivery mediated by synthetic dendritic peptide"Proceedings of the conference on frontiers Drug development. 57-58 (1999)

Norio Hazemoto 等人：“合成树突肽介导的基因传递”前沿药物开发会议记录。

Masaki Tokunaga et al.: "Effect of oligopepticles on gene expression : comparison of DNA/peptide and DNA/peptide/liposome complexes"International Journal of Phormaceutics. 269. 71-80 (2004)

Masaki Tokunaga 等人：“寡肽对基因表达的影响：DNA/肽和 DNA/肽/脂质体复合物的比较”国际药剂学杂志。

Norio Hazemoto: "Gene delivery mediated by synthetic dendritic peptide"Proceedings of the Conference on Frontiers Drug Development. 57-58 (1999)

Norio Hazemoto：“合成树突肽介导的基因传递”前沿药物开发会议记录。

Masaki Tokunaga et al.: "DNA Transfection Mediated by Synthetic Polycationic Peptides"J.Pharm.Sci.Technol.Japan. 63. 71-78 (2003)

Masaki Tokunaga等：“合成聚阳离子肽介导的DNA转染”J.Pharm.Sci.Technol.Japan。

M.Tokunaga, M.Nagao, T.M.Nagata, N.Hazemoto, T.Yotsuyanagi: "DNA Transfection Mediated by Synthetic, Polvcationic Peptides"J.Pharm.Sci.Technol., Jpn. Vol.63, No.2. 71-78 (2003)

M.Tokunaga、M.Nagao、T.M.Nagata、N.Hazemoto、T.Yotsuyanagi：“合成多阳离子肽介导的 DNA 转染”J.Pharm.Sci.Technol.，Jpn.