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1. 发明授权

US5739018A Packaging cell lines for pseudotyped retroviral vectors 失效
标题翻译：包装用于假型逆转录病毒载体的细胞系
公开(公告)号：US5739018A
公开(公告)日：1998-04-14
申请号：US694652
申请日：1996-08-07
申请人： Atsushi Miyanohara , Jiing-Kuan Yee , Shin-Tai Chen , Charles Edward Prussak , Theodore Friedmann
发明人： Atsushi Miyanohara , Jiing-Kuan Yee , Shin-Tai Chen , Charles Edward Prussak , Theodore Friedmann
IPC分类号： C12N15/63 , C12N15/867 , C12N5/10 , C12N15/86
CPC分类号： C12N15/86 , C12N15/635 , C12N2740/13052 , C12N2830/002 , C12N2830/006 , C12N2830/15 , C12N2830/30 , C12N2840/20 , C12N2840/203
摘要： The present invention features packaging cell lines and recombinant retroviral particles produced thereby, particularly pseudotyped retroviral particles. Preferably, the packaging cell lines are derived from HeLa, Cf2Th, D17, MDCK, or BHK cells, most preferably from Cf2Th cells. Retroviral particles are produced by inducibly expressing an envelope protein of interest (e.g., a retroviral envelope or the envelope protein of vesicular stomatitis virus (VSV G)). Inducible expression of the envelope protein is accomplished by operably linking an envelope protein-encoding nucleotide sequence to an inducible promoter (e.g., a promoter composed of a minimal promoter linked to multiple copies of tetO, the binding site for the tetracycline repressor (tetR) of the Escherichia coli, tetracycline resistance operon Tn10). Expression from the inducible promoter is regulated by a multi-chimeric transactivating factor, composed of a first ligand-binding domain that negatively regulates transcription from the inducible promoter (e.g., a prokaryotic tetracycline repressor polypeptide (tetR)), a transcriptional activation domain, and a second ligand-binding domain (e.g., a ligand-binding domain of a steroid receptor, preferably an estrogen receptor (ER)).
摘要翻译：本发明的特征在于包装细胞系和由此产生的重组逆转录病毒颗粒，特别是假型逆转录病毒颗粒。优选地，包装细胞系衍生自HeLa，Cf2Th，D17，MDCK或BHK细胞，最优选来自Cf2Th细胞。逆转录病毒颗粒通过诱导表达感兴趣的包膜蛋白（例如，逆转录病毒包膜或水泡性口炎病毒的包膜蛋白（VSV G））产生。通过将包膜蛋白编码核苷酸序列可操作地连接到诱导型启动子（例如，由与tetO的多拷贝连接的最小启动子，四环素阻遏物（tetR）的结合位点）组成的启动子来实现包膜蛋白的诱导表达大肠杆菌，四环素抗性操纵子Tn10）。来自诱导型启动子的表达由多嵌合反式激活因子调节，所述多嵌合反式激活因子由负调节来自诱导型启动子的转录的第一配体结合结构域（例如，原核四环素阻遏物多肽（tetR）），转录激活结构域和第二配体结合结构域（例如，类固醇受体的配体结合结构域，优选雌激素受体（ER））。

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