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    GENE TRAP CASSETTES FOR RANDOM AND TARGETED CONDITIONAL GENE INACTIVATION
    2.
    发明授权
    GENE TRAP CASSETTES FOR RANDOM AND TARGETED CONDITIONAL GENE INACTIVATION 有权
    基因诱捕技术用于随机和有针对性条件基因失活的研究

    公开(公告)号：EP1815001B8

    公开(公告)日：2011-03-23

    申请号：EP05813398.4

    申请日：2005-11-28

    申请人： Helmholtz Zentrum München Deutsches Forschungszentrum für Gesundheit und Umwelt (GmbH) ,  MPG Max-Planck-Gesellschaft zur Förderung der Wissenschaften e.V.

    发明人： VON MELCHNER, Harald ,  SCHNÜTGEN, Frank ,  WURST, Wolfgang ,  RUIZ, Patricia ,  DE-ZOLT, Silke ,  FLOSS, Thomas ,  HANSEN, Jens

    IPC分类号： C12N15/90 ,  A01K67/027 ,  C12N5/10

    CPC分类号： C12N15/1051 ,  C12N15/907 ,  C12N2799/027 ,  C12N2800/30 ,  C12N2800/60

    摘要： The present invention provides for a new type of gene trap cassettes, which can induce conditional mutations. The cassettes rely on directional site-specific recombination systems, which can repair and re-induce gene trap mutations when activated in succession. After the gene trap cassettes are inserted into the genome of the target organism, mutations can be activated at a particular time and place in somatic cells. In addition to their conditional features, the gene trap cassettes create multipurpose alleles amenable to a wide range of post-insertional modifications. Such gene trap cassettes can be used to mutationally inactivate all cellular genes. In addition the invention relates to a cell, preferably a mammalian cell which contains the above mentioned gene trap cassette. Moreover, the invention relates to the use of said cell for identification and/or isolation of genes and for the creation of transgenic organisms to study gene function at various developmental stages, including the adult, as well as for the creation of animal models of human disease useful for in vivo drug target validation. In conclusion, the present invention provides a process which enables a temporally and/or spatially restricted inactivation of all genes that constitute a living organism.

    摘要翻译： 本发明提供了一种新型的基因捕获盒，其可以诱导条件性突变。 盒依赖于定向位点特异性重组系统，当连续激活时，它可以修复并重新诱导基因陷阱突变。 在将基因捕获盒插入到靶生物体的基因组中后，可以在特定时间将突变激活并置于体细胞中。 除了它们的条件特征之外，基因捕获盒还创建了适用于广泛的插入后修饰的多用途等位基因。 这种基因捕获盒可用于突变灭活所有细胞基因。 此外，本发明涉及细胞，优选含有上述基因捕获盒的哺乳动物细胞。 此外，本发明涉及所述细胞用于基因的鉴定和/或分离以及用于创建转基因生物以研究包括成人在内的各种发育阶段的基因功能以及用于创建人的动物模型 用于体内药物靶标验证的疾病。 总之，本发明提供了一种方法，其能够在时间和/或空间上限制组成活的生物体的所有基因的失活。

    VECTORS AND METHODS FOR GENERATING VECTOR-FREE INDUCED PLURIPOTENT STEM (IPS) CELLS USING SITE-SPECIFIC RECOMBINATION
    5.
    发明公开
    VECTORS AND METHODS FOR GENERATING VECTOR-FREE INDUCED PLURIPOTENT STEM (IPS) CELLS USING SITE-SPECIFIC RECOMBINATION 审中-公开
    于产生矢量免费诱导多能干细胞的载体和方法（iPS细胞）使用SET特异性重组

    公开(公告)号：EP2268796A1

    公开(公告)日：2011-01-05

    申请号：EP09722024.8

    申请日：2009-03-17

    申请人： Helmholtz Zentrum München Deutsches Forschungszentrum für Gesundheit und Umwelt (GmbH)

    发明人： KÜHN, Ralf ,  WURST, Wolfgang

    IPC分类号： C12N5/06 ,  C12N15/09

    CPC分类号： C07K14/4702 ,  C12N5/0696 ,  C12N15/85 ,  C12N2501/602 ,  C12N2501/603 ,  C12N2501/604 ,  C12N2501/606 ,  C12N2510/00 ,  C12N2800/30 ,  C12N2830/003 ,  C12N2840/206

    摘要： The present invention relates to a DNA molecule comprising: (a) a first DNA sequence comprising: (aa) a coding sequence giving rise upon transcription to a factor that contributes to the reprogramming of a somatic cell into an induced pluripotent stem (iPS) cell; (ab) a promoter mediating the transcription of said coding sequence; and (ac) two sequence motifs that mediate excision of (aa) and/or (ab) from the DNA molecule, wherein one sequence motif is positioned 5′ and the other sequence motif is positioned 3′ of the sequence to be excised; (b) a second DNA sequence comprising a sequence motif that mediates site-specific integration of (a) into another DNA molecule. Further, the invention relates to DNA molecule comprising: (a) a first DNA sequence comprising: (aa) a coding sequence giving rise upon transcription to a factor that contributes to the reprogramming of a somatic cell into an induced pluripotent stem cell; and (ab) a promoter mediating the transcription of said coding sequence; (b) a second DNA sequence comprising: (ba) a sequence motif that mediates extrachromosomal self-replication of the DNA-molecule; and (bb) two sequence motifs that mediate excision of at least said sequence motif of (ba) from the second DNA sequence (b), wherein one sequence motif is located 5′ of (ba) and the other sequence motif 3′ of (ba). Also, the invention relates to a vector comprising the DNA molecule of the invention, a method for assembly of said vector and a somatic cell comprising said DNA molecule or said vector of the invention. Furthermore, the invention relates to methods to generate an induced pluripotent stem (iPS) cell, an induced pluripotent stem cell obtainable by said methods, to a kit comprising the DNA molecule of the invention, to a cell line or cell culture collection comprising the induced pluripotent stem cell of the invention, to the use of said cell or cell line as a research tool, to a method to generate a transgenic non-human animal and to a non-human animal generated by said method. Finally, the invention relates to a composition for gene therapy, regenerative medicine, cell therapy or drug screening.

    METHOD TO IDENTIFY MODULATORS OF B-RAF PROTEIN KINASE AND THEIR USE FOR THE TREATMENT OF ANXIETY AND DEPRESSION
    6.
    发明公开
    METHOD TO IDENTIFY MODULATORS OF B-RAF PROTEIN KINASE AND THEIR USE FOR THE TREATMENT OF ANXIETY AND DEPRESSION 审中-公开
    方法用于识别B-RAF激酶蛋白和调节剂及其用途抑郁和焦虑状态的治疗

    公开(公告)号：EP2162746A2

    公开(公告)日：2010-03-17

    申请号：EP08758979.2

    申请日：2008-06-03

    申请人： Helmholtz Zentrum München Deutsches Forschungszentrum für Gesundheit und Umwelt (GmbH)

    发明人： HITZ, Christiane ,  HÖLTER, Sabine ,  KÜHN, Ralf ,  WURST, Wolfgang ,  WEFERS, Benedikt

    IPC分类号： G01N33/68 ,  C12Q1/48

    CPC分类号： C12Q1/485 ,  C12Q1/6883 ,  C12Q2600/136 ,  C12Q2600/158 ,  G01N33/6893 ,  G01N2500/02 ,  G01N2500/10 ,  G01N2800/301 ,  G01N2800/304

    摘要： The present invention relates to a method for identifying a compound capable of modulating an anxiety or depression disorder comprising the steps of: (a) contacting a composition comprising a B-Raf protein or a B-Raf gene in expressible form or a transcript thereof with a compound under conditions that allow for an interaction of the B-Raf protein or the B-Raf gene or a transcript thereof and the compound; and (b) measuring whether said interaction, if any, results in (i) a change of B-Raf kinase activity compared to B-Raf kinase activity in the absence of said compound; (ii) a modulation of the expression of the B-Raf gene compared to B-Raf gene expression in the absence of said compound; or (iii) the formation of a complex between the compound and the B-Raf protein, wherein such a change in activity, modulation of expression or the formation of a complex is indicative of the compound being a modulator of an anxiety or depression disorder. Further, the invention relates to a method for treating an anxiety or depression disorder in an individual comprising administering to the individual an effective amount of a compound inhibiting B-Raf kinase activity or gene expression and to a use of a compound that inhibits B-Raf kinase activity or gene expression in the manufacture of a pharmaceutical composition for treating an anxiety or depression disorder. Moreover, the invention relates to a method of diagnosing a B-Raf associated anxiety or depression disorder and to a genetically engineered mouse. Finally, the invention also relates to a method of identifying another gene contributing to the pathophysiology of an anxiety or depression disorder apart from B-Raf.

    IMPROVED RECOMBINATION EFFICIENCY BY INHIBITION OF NHEJ DNA REPAIR
    9.
    发明公开
    IMPROVED RECOMBINATION EFFICIENCY BY INHIBITION OF NHEJ DNA REPAIR 审中-公开
    VERBESSERTE REKOMBINATIONSEFFIZIENZ DURCH HEMMUNG DER NHEJ-DNA-REPARATUR

    公开(公告)号：EP2718446A2

    公开(公告)日：2014-04-16

    申请号：EP12726433.1

    申请日：2012-06-06

    申请人： Helmholtz Zentrum München Deutsches Forschungszentrum für Gesundheit und Umwelt (GmbH)

    发明人： KÜHN, Ralf ,  WURST, Wolfgang

    IPC分类号： C12N15/90

    CPC分类号： C12N15/85 ,  C12N15/1024 ,  C12N15/8509 ,  C12N15/907

    摘要： The present invention relates to a method for modifying a target sequence in the genome of a mammalian cell, the method comprising the step of introducing into a mammalian cell: a. one or more compounds that introduce double-strand breaks in said target sequence; b. one or more DNA molecules comprising a donor DNA sequence to be incorporated by homologous recombination into the genomic DNA of said mammalian cell within said target sequence, wherein said donor DNA sequence is flanked upstream by a first flanking element and downstream by a second flanking element, wherein said first and second flanking element are different and wherein each of said first and second flanking sequence are homologous to a continuous DNA sequence on either side of the double-strand break introduced by said one or more compounds of a. within said target sequence in the genome of said mammalian cell; and c. one or more compounds that decrease the activity of the non-homologous end joining (NHEJ) DNA repair complex in said mammalian cell. Further, the invention relates to a method of producing a non-human mammal carrying a modified target sequence in its genome.

    摘要翻译： 本发明涉及一种用于修饰哺乳动物细胞基因组中的靶序列的方法，所述方法包括引入哺乳动物细胞的步骤：a。 一种或多种在所述靶序列中引入双链断裂的化合物; 湾 一个或多个DNA分子，其包含通过同源重组并入所述靶序列内的所述哺乳动物细胞的基因组DNA中的供体DNA序列，其中所述供体DNA序列在第一侧翼元件的上游侧和第二侧翼元件的下游， 其中所述第一和第二侧翼元件是不同的，并且其中所述第一和第二侧翼序列中的每一个与由所述一种或多种a的化合物引入的双链断裂的任一侧上的连续DNA序列同源。 在所述哺乳动物细胞的基因组中的所述靶序列内; 和c。 一种或多种降低所述哺乳动物细胞中非同源末端连接（NHEJ）DNA修复复合物的活性的化合物。 此外，本发明涉及生产在其基因组中携带修饰的靶序列的非人哺乳动物的方法。