Development of real-time intracellular pH sensor using a membrane-permeable peptide nucleic acid.

使用膜渗透性肽核酸开发实时细胞内 pH 传感器。

• 批准号: 16500200
• 负责人: TOMIZAWA Kazuhito
• 金额: $ 2.37万
• 依托单位: Okayama University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 2004
• 资助国家: 日本
• 起止时间: 2004 至 2005
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-16500200/
• 关键词: Peptide nucleic acid; Polyarginine; Nucleic acid; Bio-molecule; Bio-imaging; Cell; 蛋白質導入法; TAT; センサー; リアルタイム; イメージング; 蛋白導入法; FRET

Membrane-permeable peptide nucleic acid (PNA) developed by us is thought to be a powerful tool for the regulation of cell function. The purpose of this project is that development of the tools that are useful for the regulation of protein expression and the function and for the monitoring of nano-ecology in living cells using the PNA. In the present project, we show that membrane-permeable PNA is a powerful tool for the regulation of gene expression and for anti-cancer therapy as follows.1.Regulation of gene expression.We synthesized a PNA, which effectively hybridized with plasmid DNA, and the PNA was fused a membrane-permeable poly-arginine to have the ability of membrane permeability. After mixing the PNA with plasmid DNAs, cells were incubated with the complex and the efficiency of the transfection was investigated. The plasmid DNAs were effectively transfected in both cells and the efficiency is better than that of cationic transfection method.2.Antigene therapy using membrane-permeable PNA.We synthesized PNAs that are able to bind with p53 mRNA and the double-strand DNA. Cancer cells were incubated with the PNAs. The PNAs were delivered into cells and inhibited the expression of p53 mRNA and the protein. The combination treatment of the PNAs with p53 protein therapy was capable to replace endogenous mutated p53 proteins with wild-type p53 proteins in the cancer cells.

我们开发的膜透性肽核酸（PNA）被认为是调节细胞功能的有力工具。该项目的目的是开发用于调节蛋白质表达和功能以及使用PNA监测活细胞中纳米生态的工具。本课题的主要研究内容如下：1.基因表达调控：合成了一种能与质粒DNA有效杂交的肽核酸，并将其与具有膜通透性的聚精氨酸融合，使其具有膜通透性。将PNA与质粒DNA混合后，将细胞与复合物一起孵育，并研究转染效率。质粒DNA在两种细胞中均能有效转染，且转染效率优于阳离子转染法。2.膜渗透性PNA的抗基因治疗：我们合成了能与p53 mRNA和双链DNA结合的PNA。将癌细胞与PNA一起孵育。PNAs可被导入细胞内，抑制p53 mRNA和蛋白的表达。PNA与p53蛋白治疗的组合治疗能够在癌细胞中用野生型p53蛋白替代内源性突变的p53蛋白。

项目成果

p53 protein transduction therapy: Successful targeting and inhibition of the growth of the bladder cancer cells

• DOI: 10.1016/j.eururo.2005.08.019
• 发表时间: 2006-01-01
• 期刊: EUROPEAN UROLOGY
• 影响因子: 23.4
• 作者: Inoue, M;Tomizawa, K;Matsui, H
• 通讯作者: Matsui, H

タンパク質セラピー法

蛋白质疗法

• 发表时间: 2004
• 期刊: 実験医学 Vol.22・No.18
• 作者: 松下正之;富澤一仁;松井秀樹
• 通讯作者: 松井秀樹

Regulation of synaptic vesicle recycling by calcineurin in different vesicle pools

• DOI: 10.1016/j.neures.2004.12.018
• 发表时间: 2005-04-01
• 期刊: NEUROSCIENCE RESEARCH
• 影响因子: 2.9
• 作者: Kumashiro, S;Lu, YF;Matsui, H
• 通讯作者: Matsui, H

The NH2-terminal of influenza virus hemagglutinin-2 subunit peptides enhance the anti-tumor potency of poly-arginine-mediated p53 protein transduction.

流感病毒血凝素 2 亚基肽的 NH2 末端增强聚精氨酸介导的 p53 蛋白转导的抗肿瘤效力。

• 发表时间: 2005
• 期刊: J.Biol.Chem. 280(9)
• 作者: Michiue H;Tomizawa K;Wei FY;Matsushita M;Lu YF;Ichikawa T;Tamiya T;Date I;Matsui H.
• 通讯作者: Matsui H.

Control of cyclin‐dependent kinase 5 (Cdk5) activity by glutamatergic regulation of p35 stability

• DOI: 10.1111/j.1471-4159.2005.03058.x
• 发表时间: 2005-04
• 期刊: Journal of Neurochemistry
• 影响因子: 4.7
• 作者: Fan-Yan Wei;K. Tomizawa;T. Ohshima;A. Asada;Taro Saito;C. Nguyen;J. Bibb;K. Ishiguro;Ashok B Kulkarni;H. Pant;K. Mikoshiba;H. Matsui;S. Hisanaga