Knock-down mouse, a novel gene engineering mouse for use as a disease model

击倒小鼠，一种用作疾病模型的新型基因工程小鼠

• 批准号: 11480251
• 负责人: OHNISHI Yasuyuki
• 金额: $ 9.41万
• 依托单位: Central Institute for Experimental Animals
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (B).
• 财政年份: 1999
• 资助国家: 日本
• 起止时间: 1999 至 2000
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-11480251/
• 关键词: ribozyme; mouse; insulin; vascular endotherial growth factor; xenotransplantability; K-ras; human neoplasm; tetracyclin; スイッチ; トランスジェニック

In this research, we attempt to create a novel gene engineering mouse which contains the target gene expression-switch such as tetracycline regulated gene expression system (Tet-OFF and Tet-ON) and a gene for ribizyme driven by the switch system. First, we constructed an anti-insulin ribozyme, and examined in vitro cleaving assay using synthesized insulin RNA and the ribozyme. The ribozyme showed in vitro cleavage activity to the RNA.The ribozyme also showed anti-insulin activity in M6 insulinoma cell transfectant system. Another ribozymes were created for VEGF (vascular endotherial growth factor) and activated K-ras gene RNA, and they showed cleaving activity in cell transformant and in vitro cleavage assay system.Also, tumor cells transfected by the anti-VEGF or anti-K-ras ribozyme showed reduced tumorigenic activity in nude mouse. Transgenic mice for rusiferase gene driven by the Tet-OFF and Tet-ON system were created. The mice were under exanimation.

在本研究中，我们试图创建一种新的基因工程小鼠，该小鼠包含目标基因表达开关，如四环素调控的基因表达系统（Tet-OFF和Tet-ON）和由该开关系统驱动的ribizyme基因。首先，我们构建了一种抗胰岛素的核酶，并利用合成的胰岛素RNA和核酶进行了体外切割实验。该核酶在体外具有切割RNA的活性，在M6胰岛素瘤细胞转染系统中也具有抗胰岛素活性。另一种针对VEGF（vascular endotherial growth factor，血管内皮生长因子）和活化的K-ras基因RNA的核酶，在细胞培养和体外切割系统中显示出切割活性，转染抗VEGF或抗K-ras核酶的肿瘤细胞在裸鼠体内的致瘤活性也有所降低。建立了由Tet-OFF和Tet-ON系统驱动的转rusiferase基因小鼠。小鼠处于兴奋状态。

项目成果

Okamoto, K., Oshika, K., Fukushima, Y., Ohnishi, Y., Tokunaga, T., Tomii, Y., Kijima, H., Yamazaki H., Ueyama, Y., Tamaoki, N.and Nakamura, M.: "Xenografts of solid tumors frequently express cellular-associated isoform of vascular endothelial growth facto

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Ueyama, Y., Abe, Y., Ohnishi, Y., Sawa, N., Hatanaka, H., Handa, A., Tokuda, Y., Yamazaki, H., Kijima, H., Tamaoki, N.and Nakamura, M.: "In vivo chemosensitivity of human malignant cystosarcoma phyllodes xenograft."Oncol.Rep.. 7. 257-260 (2000)

上山 Y.、阿部 Y.、大西 Y.、泽 N.、畑中 H.、半田 A.、德田 Y.、山崎 H.、木岛 H.、玉木 N. 和

Oshika,Y: "Ribozyme approach to downregulate vascular endothelial growth factor (VEGF) 189 expression in non-small cell lung cancer (NSCLC)"Eur.J.Cancer. 36. 2390-2396 (2000)

Oshika,Y：“核酶方法下调非小细胞肺癌 (NSCLC) 中血管内皮生长因子 (VEGF) 189 的表达”Eur.J.Cancer。

Ohnishi, Y., Arai, T., Koshirakawa, M., Horii, N., Nakajo, S., Urano, K., Usui, T., Tamaoki, N., and Ueyama, Y.: "Induction of drug metabolism-related enzymes by methylcholanthrene and phenobarbital in transgenic mice carrieng human prototype c-Ha-ras gen

Ohnishi, Y.、Arai, T.、Koshirakawa, M.、Horii, N.、Nakajo, S.、Urano, K.、Usui, T.、Tamaoki, N. 和 Ueyama, Y.：“药物的诱导

Tokunaga,T.: "Ribozyme-mediated inactivation of mutant K-ras oncogene in a colon cancer cell line"Br.J.Cancer. 83. 833-839 (2000)

Tokunaga,T.：“核酶介导的结肠癌细胞系中突变 K-ras 癌基因的失活”Br.J.Cancer。