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Generation and analysis of transgenic mice carrying parvoviral NS gene

携带细小病毒NS基因的转基因小鼠的产生和分析

基本信息

批准号：
08680903
负责人：
YAGAMI Ken-ichi
金额：
$ 1.54万
依托单位：
University of Tsukuba
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (C)
财政年份：
1996
资助国家：
日本
起止时间：
1996 至 1997
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-08680903/
关键词：
Parvovirus Transgenic mice Nonstractural protein Tet-inducible system Cytotoxicity 病原性

项目摘要

Parvoviruses have small single stranded DNA,its nonstructural (NS) protein regulates viral capsid protein (VP) and host gene expressions. Correlation between parvoviruses and autoimmun diseases or antitumorigenesis suspected because of modification of immunocompetence. Thus, we tried to generate transgenic mice to involve modification of autoimmune disease and anti tumorigenesis with parvoviral NS protein.At first, tetracycline inducible gene expression system was applied to generate transgenic mice overexperssing prvoviral NS,because of potential cytotoxicity of NS to mouse fetus. We generated 4 founder mice carrying reverse tetracycline transactivator (rtTA) gene and 3 founder mice carrying fusion transgene including tetracycline responsive element (TRE) and parvoviral NS genes. These mice are cross breeding to generate dual transgenic mice overexpression NS by administration of tetracycline.On the other hand, we also examined cytotoxicity of NS and cellular factors interact with NS in host cells, to expect phenotypes by overexpression of NS.The results indicate that cytotoxicity of NS is correlated with induction of apoptosis, and that NS interacts to CBP (CREB binding protein) which is an important coactivators to bind several transcriptional factors.

细小病毒具有小的单链DNA，其非结构（NS）蛋白调节病毒衣壳蛋白（VP）和宿主基因表达。由于免疫能力的改变，怀疑细小病毒与自身免疫性疾病或抗肿瘤发生之间的相关性。因此，我们试图用细小病毒NS蛋白来产生转基因小鼠，以涉及自身免疫性疾病的修饰和抗肿瘤发生。首先，由于NS对小鼠胎儿具有潜在的细胞毒性，应用四环素诱导基因表达系统来产生过度表达细小病毒NS的转基因小鼠。我们产生了 4 只携带反向四环素反式激活子 (rtTA) 基因的创始小鼠和 3 只携带融合转基因的创始小鼠，其中融合转基因包括四环素反应元件 (TRE) 和细小病毒 NS 基因。这些小鼠通过四环素杂交产生过表达NS的双转基因小鼠。另一方面，我们还检测了NS的细胞毒性以及宿主细胞中与NS相互作用的细胞因子，以预期NS过表达的表型。结果表明NS的细胞毒性与诱导细胞凋亡相关，并且NS与CBP相互作用（CREB结合蛋白），它是结合多种转录因子的重要共激活剂。