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生体認識分子を融合したナノ粒子による全く新しい生体内ピンポイント遺伝子導入法

使用与生物识别分子融合的纳米颗粒的全新体内精确基因转移方法

基本信息

批准号：
13218080
负责人：
黒田俊一
金额：
$ 3.9万
依托单位：
Osaka University
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research on Priority Areas (C)
财政年份：
2001
资助国家：
日本
起止时间：
2001 至无数据
项目状态：
已结题

项目摘要

本研究では、高い細胞及び組織特異性を有し、極めて高い遺伝子導入効率を有するウイルス感染機構を応用し、かつ、ウイルスゲノムを完全に排除した副作用の危険性が低い遺伝子導入法を開発することを目的としている。(1)組換え酵母によるHBsAg (B型肝炎ウイルス(HBV)表面抗原(HBsAg)粒子)の大量生産:ヒト肝臓に対する特異的レセプターを有するHBsAg Lタンパク質粒子を酵母で大量に生産する方法を確立した。(2)同粒子を用いるヒト肝臓癌由来培養細胞への遺伝子導入:エレクトロポレーション法により粒子内部にGFP(緑色蛍光タンパク質)発現遺伝子を封入し、ヒト肝癌由来細胞HepG2に添加したところ、リポフェクタミン法よりも百倍以上高い効率で、ほぼ全ての細胞に遺伝子を導入した。(3)同粒子を用いるヒト肝臓癌を接種したヌードマウスへの遺伝子導入:ヒト肝臓癌由来細胞HuH-7とヒト大腸癌由来細胞WiDrを接種したヌードマウスにGFP発現ベクターを含む同粒子を腹腔内投与したところ、ヒト肝臓癌由来固形癌部にGFPによる蛍光が認められた。一方、マウスの通常臓器、及びヒト大腸癌由来固形癌部には全く蛍光は認められなかった。(4)同粒子を用いるヒト肝臓癌を接種したヌードマウスにおける遺伝子治療:最近では、本粒子内部にHSV由来チミジンキナーゼ遺伝子を包含し、ヒト肝臓癌を移植したヌードマウスに静脈注射し、ガンシクロビルを経口投与することで、移植ヒト肝癌特異的な増殖抑制効果を観察することができた。以上の結果から、本粒子を用いた遺伝子導入方法は、高い細胞及び組織特異性を有し、極めて高い遺伝子導入効率を有するウイルス感染機構を応用し、かつ、ウイルスゲノムを完全に排除した副作用の危険性が低いと考えられた。

In this study, there are some characteristics in this study, such as high cell, high cell and tissue characteristics, and extremely high enrollment rates. in this study, the incidence of infection was significantly higher than that of normal controls in this study. in this study, we completely excluded the side effects, dangerous, dangerous, low-risk, low-risk, low-risk (1) A large number of HBsAg (hepatitis B virus surface antigen (HBV) surface antigen (HBsAg) particles) were produced in large quantities. There was a large amount of HBsAg L particles in the liver. (2) the same particles were used to detect the origin of liver cancer. The cells were divided into the same particle, the GFP (color light), the cell HepG2, the cancer cell, the cell. (3) the same particles were used to transfer the same particles into the liver cancer cells. (3) the same particles were inoculated into HuH-7 cancer cells. (3) the same particles were injected into the abdominal cavity of liver cancer cells. (3) the same particles were injected into the abdominal cavity of liver cancer. (3) the same particles were injected into the abdominal cavity of liver cancer. (3) the same particles were injected into the abdominal cavity of liver cancer. (3) the same particles were injected into the abdominal cavity of liver cancer. (3) the same particles were injected into the abdominal cavity of liver cancer. (3) the same particles were injected into the abdominal cavity of liver cancer. (3) the same particles were injected into the abdominal cavity of liver cancer. (3) the same particles were injected into the abdominal cavity with the same particles. (3) the same particles were injected into the abdominal cavity of the liver cancer. The origin of the liver cancer is the origin of HuH-7 cancer, the origin of the liver cancer, the origin of the liver cancer, the origin of the cancer, the One side, the usual device, and the large part of the solid cancer is caused by total light exposure. (4) the same particle was used to treat liver cancer. Recently, the origin of the HSV in this particle was used to treat liver cancer. This is the cause of the disease. Transplantation of hepatoma-specific colonization inhibition results were observed. The results of the above results showed that the results of the above results showed that there were significant differences in the results of the above results, the results of the above results