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    • 7. 发明授权
    • Methods for creating and identifying functional RNA interference elements
    • 用于创建和鉴定功能性RNA干扰元件的方法
    • US08735064B2
    • 2014-05-27
    • US12735222
    • 2008-12-23
    • David MicklemJames Lorens
    • David MicklemJames Lorens
    • C12P19/34C12Q1/68C07H21/02
    • C12N15/1086C12N15/10C12N15/111C12N15/64C12N15/66C12N2310/14C12N2320/10C12N2330/30C12Q1/6897
    • The invention relates to the control of gene expression. Specifically, the invention provides compositions and methods for the production and use of recombinant nucleic acid molecules that have the ability to specifically downregulate an expressed target gene in vivo. In some aspects, the invention provides methods for producing a hairpin DNA molecule where part of the molecule is derived from an mRNA that is a target for a small interfering RNA (siRNA) derived from the hairpin. In other aspects, the invention provides synthetic hairpin adapter oligonucleotides that are used in the construction of siRNA-producing cassettes. In other aspects, the invention provides methods for testing for the presence or absence of specific inhibitory activity of an RNAi trigger molecule, and in still other aspects, the invention provides methods for identifying an active RNAi trigger molecule from a library of RNAi trigger molecules. In still other aspects, the invention provides methods for identifying a polynucleotide from a plurality of candidate target polynucleotides that is specifically targeted by an RNAi trigger molecule. In other aspects, the invention provides epi-allelic series of hypomorphic RNAi trigger molecules specific for any gene of interest, where the series of RNAi trigger molecules have a variety of uses including analysis of gene function and drug target development.
    • 本发明涉及基因表达的控制。 具体而言,本发明提供了生产和使用具有在体内特异性下调表达的靶基因的能力的重组核酸分子的组合物和方法。 在一些方面,本发明提供了用于产生发夹DNA分子的方法,其中分子的一部分衍生自作为衍生自发夹的小干扰RNA(siRNA)的靶的mRNA。 在其它方面,本发明提供用于构建产siRNA盒的合成发夹衔接寡核苷酸。 在其他方面,本发明提供了用于测试RNAi触发分子的特异性抑制活性的存在或不存在的方法,并且在其它方面,本发明提供了从RNAi触发分子文库鉴定活性RNAi触发分子的方法。 在其它方面,本发明提供了从由RNAi触发分子专门靶向的多个候选靶多核苷酸中鉴定多核苷酸的方法。 在其它方面,本发明提供了对于任何目的基因特异性的异型RNAi触发分子的等位基因系列,其中一系列RNAi触发分子具有各种用途,包括基因功能分析和药物靶标开发。
    • 10. 发明授权
    • Shuttle vectors
    • 班车向量
    • US06391582B2
    • 2002-05-21
    • US09208827
    • 1998-12-09
    • Ying LuoPei Wen YuJames Lorens
    • Ying LuoPei Wen YuJames Lorens
    • C12P2102
    • C12N15/1082C12N15/1034C12N15/70C12N15/81
    • The invention provides shuttle vectors, and methods of using shuttle vectors, capable of expression in, at least, a mammalian cell. Furthermore, the shuttle vectors are capable of replication in at least yeast, and optionally, bacterial cells. Also provided is a method wherein yeast are transformed with a shuttle vector as provided herein. Heterologous nucleic acids flanked by 5′ and 3′ ends identical to a homologous recombination site within the shuttle vector are introduced to the transformed yeast and allowed to homologously recombine with the shuttle vector such that they are inserted into the vector by the yeast organism. The shuttle vector is then recovered and transferred to a mammalian cell for expression.
    • 本发明提供穿梭载体,以及使用能够在至少哺乳动物细胞中表达的穿梭载体的方法。 此外,穿梭载体能够在至少酵母和任选的细菌细胞中复制。 还提供了一种方法,其中用本文提供的穿梭载体转化酵母。 将与穿梭载体内的同源重组位点相同的5'和3'末端的异源核酸引入转化的酵母中,并与穿梭载体同源重组,使得它们被酵母生物插入载体。 然后将穿梭载体回收并转移至哺乳动物细胞进行表达。