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    • 3. 发明授权
    • Compositions and methods for helper-free production of recombinant adeno-associated viruses
    • 无辅助生产重组腺相关病毒的组合物和方法
    • US06258595B1
    • 2001-07-10
    • US09404555
    • 1999-09-23
    • Guang-Ping GaoJames M. Wilson
    • Guang-Ping GaoJames M. Wilson
    • C12N1586
    • G01R31/275C12N15/86C12N2750/14143C12N2750/14151C12N2840/20G01R31/013G01R31/316
    • A method for producing recombinant adeno-associated virus in the absence of contaminating helper virus or wild-type virus involves culturing a mammalian host cell containing an rAd/AAV hybrid virus, an AAV rep sequence and an AAV cap sequence under the control of regulatory sequences directing expression thereof. The rAd/AAV hybrid virus contains a rAAV construct to be packaged into an AAV virion in a backbone containing the adenoviral sequences necessary to express E1a and E1b gene products and to permit replication of the hybrid virus. The method of the invention permits replication of the hybrid virus and production of rAAV virion in this host cell in the absence of a helper virus and obviates a subsequent purification step to purify rAAV from contaminating virus.
    • 在不存在污染性辅助病毒或野生型病毒的情况下用于产生重组腺相关病毒的方法包括在调节序列的控制下培养含有rAd / AAV杂合病毒,AAV rep序列和AAV帽序列的哺乳动物宿主细胞 指导其表达。 rAd / AAV杂合病毒含有一个rAAV构建体,其被包装在含有表达E1a和E1b基因产物所必需的腺病毒序列并且允许杂交病毒复制的骨架中的AAV病毒粒子中。 本发明的方法允许在不存在辅助病毒的情况下复制该杂种病毒并在该宿主细胞中产生rAAV病毒粒子,并且消除随后的纯化步骤以从污染病毒纯化rAAV。