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    High throughput methods for functionally determining RNA interference efficiency
    1.
    发明授权
    High throughput methods for functionally determining RNA interference efficiency 有权
    用于功能测定RNA干扰效率的高通量方法

    公开(公告)号：US08901288B2

    公开(公告)日：2014-12-02

    申请号：US12739382

    申请日：2008-10-24

    申请人： Christof Fellmann ,  Scott W. Lowe ,  Gregory J. Hannon ,  Johannes E. Zuber

    发明人： Christof Fellmann ,  Scott W. Lowe ,  Gregory J. Hannon ,  Johannes E. Zuber

    IPC分类号： C07H21/00 ,  C12Q1/68 ,  C12N15/10

    CPC分类号： C12Q1/6897 ,  C12N15/1086 ,  C12N15/113 ,  C12N15/85 ,  C12N2310/14

    摘要： Provided is a single construct combining a sequence encoding an RNAi molecule, a sequence encoding a reporter, and a target sequence specific for the RNAi molecule. The construct can be used to determine the potency of the encoded RNAi molecule in a direct and unbiased way. These results can be used to inform the design of potent RNAi molecules of various types and can be extended to several other applications, including: (1) generation of tiled libraries comprising every possible RNAi molecule-encoding sequence for a given gene target; (2) large-scale parallel validation of RNAi molecules targeting many genes to generate validated RNAi molecule-encoding libraries; (3) experimental comparison of design algorithms and strategies; and (4) investigation of RNAi biology in target site mutagenesis assays by screening pools containing single nucleotide changes in target sites and/or in the RNAi molecule to identify the most relevant sequence characteristics of potent RNAi-target site predictions.

    摘要翻译： 提供了组合编码RNAi分子的序列，编码报道分子的序列和对RNAi分子特异的靶序列的单一构建体。 该构建体可以用于以直接和公正的方式确定编码的RNAi分子的效力。 这些结果可用于通知各种有效的RNAi分子的设计，并可扩展到其他几个应用，包括：（1）生成包含给定基因靶标的每个可能的RNAi分子编码序列的平铺文库; （2）针对许多基因的RNAi分子的大规模并行验证以产生经验证的RNAi分子编码文库; （3）设计算法和策略的实验比较; 和（4）通过在靶位点和/或RNAi分子中含有单核苷酸变化的筛选池来检测靶位点诱变测定中的RNAi生物学，以鉴定有效的RNAi靶位点预测的最相关的序列特征。

    HIGH THROUGHPUT METHODS FOR FUNCTIONALLY DETERMINING RNA INTERFERENCE EFFICIENCY
    2.
    发明申请
    HIGH THROUGHPUT METHODS FOR FUNCTIONALLY DETERMINING RNA INTERFERENCE EFFICIENCY 有权
    用于功能测定RNA干扰效率的高通量方法

    公开(公告)号：US20110015093A1

    公开(公告)日：2011-01-20

    申请号：US12739382

    申请日：2008-10-24

    申请人： Christof Fellmann ,  Scott W. Lowe ,  Gregory J. Hannon ,  Johannes Ekkehart Zuber

    发明人： Christof Fellmann ,  Scott W. Lowe ,  Gregory J. Hannon ,  Johannes Ekkehart Zuber

    IPC分类号： C40B30/06 ,  C12N15/63 ,  C40B40/06 ,  C12Q1/68 ,  C12N5/073

    CPC分类号： C12Q1/6897 ,  C12N15/1086 ,  C12N15/113 ,  C12N15/85 ,  C12N2310/14

    摘要： Provided is a single construct combining a sequence encoding an RNAi molecule, a sequence encoding a reporter, and a target sequence specific for the RNAi molecule. The construct can be used to determine the potency of the encoded RNAi molecule in a direct and unbiased way. These results can be used to inform the design of potent RNAi molecules of various types and can be extended to several other applications, including: (1) generation of tiled libraries comprising every possible RNAi molecule-encoding sequence for a given gene target; (2) large-scale parallel validation of RNAi molecules targeting many genes to generate validated RNAi molecule-encoding libraries; (3) experimental comparison of design algorithms and strategies; and (4) investigation of RNAi biology in target site mutagenesis assays by screening pools containing single nucleotide changes in target sites and/or in the RNAi molecule to identify the most relevant sequence characteristics of potent RNAi-target site predictions.

    摘要翻译： 提供了组合编码RNAi分子的序列，编码报道分子的序列和对RNAi分子特异的靶序列的单一构建体。 该构建体可以用于以直接和公正的方式确定编码的RNAi分子的效力。 这些结果可用于通知各种有效的RNAi分子的设计，并可扩展到其他几个应用，包括：（1）生成包含给定基因靶标的每个可能的RNAi分子编码序列的平铺文库; （2）针对许多基因的RNAi分子的大规模并行验证以产生经验证的RNAi分子编码文库; （3）设计算法和策略的实验比较; 和（4）通过在靶位点和/或RNAi分子中含有单核苷酸变化的筛选池来检测靶位点诱变测定中的RNAi生物学，以鉴定有效的RNAi靶位点预测的最相关的序列特征。