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    • 1. 发明申请
    • NOVEL RNA INTERFERENCE METHODS USING DNA-RNA DUPLEX CONSTRUCTS
    • 使用DNA-RNA双相结构的新型RNA干扰方法
    • WO2009049251A1
    • 2009-04-16
    • PCT/US2008/079638
    • 2008-10-10
    • UNIVERSITY OF SOUTHERN CALIFORNIA USC STEVENSYING, Shao-YaoLIN, Shi-lung
    • YING, Shao-YaoLIN, Shi-lung
    • A61K31/7088
    • C12N15/1096
    • The present invention provides novel compositions and. methods for suppressing the function or activity of a targeted gene through a novel intracellular piRNA-mediated RNAi mechanism, using RNA-DNA duplex constructs. The invention further provides novel methods and compositions for generating or producing RNA-DNA duplex agents, whose quantity is high enough to be used for the invention's gene silencing transfection and possibly in therapeutics applications. This improved RNA-polymerase chain reaction (RNA-PCR) method utilizes thermocycling steps of promoter- linked DNA or RNA template synthesis, in υitro transcription and then reverse transcription to bring up the amount of RNA-DNA duplexes up to two thousand folds within one round of the above procedure for using in D -RNAi- directed gene silencing.
    • 本发明提供新的组合物。 使用RNA-DNA双链构建体通过新的细胞内piRNA介导的RNAi机制抑制靶基因的功能或活性的方法。 本发明还提供用于产生或产生RNA-DNA双链体的新方法和组合物,其量足够高以用于本发明的基因沉默转染并且可能在治疗应用中。 这种改进的RNA聚合酶链反应(RNA-PCR)方法利用启动子连接的DNA或RNA模板合成的热循环步骤,然后逆转录将RNA-DNA双链体的量提高到2000倍以内 在D-RNAi定向基因沉默中使用上述方法的一轮。