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1. 发明申请

WO1994010329A1 RNA PACKAGING SYSTEM 审中-公开
标题翻译： RNA包装系统
公开(公告)号：WO1994010329A1
公开(公告)日：1994-05-11
申请号：PCT/US1993010396
申请日：1993-10-28
申请人： RUTGERS UNIVERSITY
发明人： RUTGERS UNIVERSITY , WILSON, Thomas, M., A. , HWANG-LEE, Duk-Ju
IPC分类号： C12P19/34
CPC分类号： C07K14/005 , C12N7/00 , C12N15/10 , C12N15/8201 , C12N15/8206 , C12N2770/00022 , Y10S435/81
摘要： The present invention relates to an in vivo system for expression and packaging of recombinant RNA into pseudovirus particles. The invention is based on the discovery that plant viral coat proteins (CPs) may be efficiently expressed in E. coli, and that these recombinant coat proteins will function to assemble in vivo and package recombinant chimeric RNA, containing an operatively linked origin-of-assembly (OAS) sequence, to form mature viral particles containing a foreign RNA. The present invention thus provides for packaging of RNA into a ribonuclease-resistant form that is easily purified and stored, and which overcomes the prior art problems associated with degradation of RNA by ribonucleases. Significantly, the method of the invention is RNA sequence- and length-independent. The components of the invention include a source in the bacterial host of viral coat proteins, and a source in the bacterial host to direct the transcription of a DNA molecule comprising an OAS-encoding DNA and a foreign DNA, which DNA molecule can be transcribed in the host cell to produce an RNA molecule comprising an OAS operatively linked to an RNA of interest. The CPs and OAS are from a plant virus having a rod-shaped helical particle and a single-stranded RNA genome, most preferably tobacco mosaic virus.
摘要翻译：本发明涉及用于将重组RNA表达和包装成假病毒颗粒的体内系统。本发明基于以下发现：可以在大肠杆菌中有效表达植物病毒外壳蛋白（CP），并且这些重组外壳蛋白将用于在体内组装并包装重组嵌合RNA，所述重组嵌合RNA含有可操作地连接的起始蛋白组装（OAS）序列，以形成含有外源RNA的成熟病毒颗粒。因此，本发明提供将RNA包装到易于纯化和储存的核糖核酸酶抗性形式中，并克服了与核糖核酸酶降解RNA相关的现有技术问题。显着地，本发明的方法是RNA序列和长度无关的。本发明的组分包括病毒外壳蛋白的细菌宿主中的源，以及细菌宿主中用于引导包含编码OAS的DNA和外来DNA的DNA分子的转录的来源，该DNA分子可被转录在宿主细胞以产生包含可操作地连接到目的RNA的OAS的RNA分子。 CP和OAS来自具有棒状螺旋颗粒和单链RNA基因组的植物病毒，最优选烟草花叶病毒。

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