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    SOMATIC HAPLOID HUMAN CELL LINE
    5.
    发明授权
    SOMATIC HAPLOID HUMAN CELL LINE 有权

    公开(公告)号：EP3074515B1

    公开(公告)日：2018-11-14

    申请号：EP14853186.6

    申请日：2014-11-28

    申请人： Horizon Discovery Limited

    发明人： BUERCKSTUEMMER, Tilmann

    IPC分类号： C12N15/11 ,  C12N15/10

    CPC分类号： C12N15/907 ,  C12N5/0602 ,  C12N15/902 ,  C12N2800/30 ,  C12Q1/6813

    摘要： The invention provides for a method of producing a mutant somatic human cell line of cells comprising a genomic mutation of interest (MOI) at a predefined genomic site of interest (GOI) in close proximity to a genomic target site, which comprises: a) providing a guide RNA (gRNA) comprising a tracrRNA in conjunction with crRNA including an oligonucleotide sequence that hybridizes with the target site; b) providing an RNA-guided endonuclease which catalyzes the DNA break at the target site upon hybridizing with the gRNA; c) introducing the gRNA into the cells in the presence of the endonuclease to obtain a repertoire of cells comprising a variety of genomic mutations at the target site; d) selecting a cell from said repertoire which comprises a MOI; wherein the cell is haploid for the genomic locus of the target site; and e) expanding the cell to obtain the mutant cell line. The invention further provides for a mutant human somatic cell line obtainable by such method; and libraries of mutant human somatic cell lines of isogenic cells with a variety of genomic mutations at different predefined genomic target sites.

    TARGETED ELIMINATION OF BACTERIAL GENES
    7.
    发明公开
    TARGETED ELIMINATION OF BACTERIAL GENES 审中-公开

    公开(公告)号：EP3224353A4

    公开(公告)日：2018-06-06

    申请号：EP15863631

    申请日：2015-11-26

    申请人： TECH INNOVATION MOMENTUM FUND ISRAEL LIMITED PARTNERSHIP

    发明人： QIMRON EHUD ,  YOSEF IDO ,  MANOR MIRIAM

    IPC分类号： C12N7/01 ,  A61K35/76 ,  A61P31/04 ,  C12N15/113 ,  C12N15/74

    CPC分类号： C12N15/902 ,  A61K35/76 ,  C12N15/111 ,  C12N15/63 ,  C12N15/74 ,  C12N2310/20 ,  C12N2320/30

    摘要： Provided is a kit or a system including two elements or components. The first component (i) is a selective component including a nucleic acid sequence and at least one proto-spacer. The second component (ii) includes at least one sensitizing component including at least one cas gene and at least one CRISPR array. At least one spacer of the CRISPR targets a proto-spacer included within a pathogenic gene of a bacterium so as to specifically inactivate said pathogenic gene in said bacterium and wherein at least one spacer of said CRISPR targets a proto-spacer included within said selective component of (i) so as to specifically inactivate said selective component. Further provided is a method using the components or kits of the invention for interference with a horizontal transfer of a pathogenic gene between bacteria and for preventing a pathologic condition in a mammalian subject caused by a bacterial infection.

    NOVEL GENE TARGETING METHOD
    8.
    发明公开
    NOVEL GENE TARGETING METHOD 审中-公开
    新型基因靶向方法

    公开(公告)号：EP3290518A1

    公开(公告)日：2018-03-07

    申请号：EP16785983.4

    申请日：2016-04-29

    申请人： Hangzhou Genekine Biotech Co., Ltd

    发明人： JIANG, Youwei ,  LI, Yunfei

    IPC分类号： C12N15/11 ,  C12N1/16 ,  C12P21/00

    CPC分类号： C12N15/905 ,  C12N1/16 ,  C12N9/1241 ,  C12N15/11 ,  C12N15/902 ,  C12N2800/30 ,  C12P21/00 ,  C12N15/102 ,  C12Q2521/507

    摘要： Provided are a novel two-step gene targeting method and a nucleotide construct for gene targeting. The method can improve the gene targeting efficiency and accurately identify a target gene knock-out mutant. The method of the present invention comprises: firstly, efficiently replacing a target gene in a genome with a targeting box by homologous recombination, the targeting box consisting of a target gene activity variant, a marker gene and site-specific recombination sites; and secondly, resecting the targeting box by recombinase, leaving a site-specific recombination site on the target gene to generate a target gene knock-out mutant, and removing a recombinase expression vector from the knock-out mutant by using a counter selection marker in the recombinase expression vector.

    摘要翻译： 提供了一种新的两步基因靶向方法和用于基因靶向的核苷酸构建体。 该方法可以提高基因靶向效率并准确鉴定靶基因敲除突变体。 本发明的方法包括：首先通过同源重组，有效置换基因组中的靶基因，所述靶向盒由靶基因活性变异体，标记基因和位点特异性重组位点组成; 其次，通过重组酶切除靶向盒，在靶基因上留下位点特异性重组位点以产生靶基因敲除突变体，并通过使用中的计数选择标记从敲除突变体中去除重组酶表达载体 重组酶表达载体。

    TARGETED ELIMINATION OF BACTERIAL GENES
    10.
    发明公开
    TARGETED ELIMINATION OF BACTERIAL GENES 审中-公开
    靶向消灭细菌基因

    公开(公告)号：EP3224353A1

    公开(公告)日：2017-10-04

    申请号：EP15863631.6

    申请日：2015-11-26

    申请人： Technology Innovation Momentum Fund (Israel) Limited Partnership

    发明人： QIMRON, Ehud ,  YOSEF, Ido ,  MANOR, Miriam

    IPC分类号： C12N7/01 ,  C12N15/113 ,  C12N15/74 ,  A61K35/76 ,  A61P31/04

    CPC分类号： C12N15/902 ,  A61K35/76 ,  C12N15/111 ,  C12N15/63 ,  C12N15/74 ,  C12N2310/20 ,  C12N2320/30

    摘要： Provided is a kit or a system including two elements or components. The first component (i) is a selective component including a nucleic acid sequence and at least one proto-spacer. The second component (ii) includes at least one sensitizing component including at least one cas gene and at least one CRISPR array. At least one spacer of the CRISPR targets a proto-spacer included within a pathogenic gene of a bacterium so as to specifically inactivate said pathogenic gene in said bacterium and wherein at least one spacer of said CRISPR targets a proto-spacer included within said selective component of (i) so as to specifically inactivate said selective component. Further provided is a method using the components or kits of the invention for interference with a horizontal transfer of a pathogenic gene between bacteria and for preventing a pathologic condition in a mammalian subject caused by a bacterial infection.

    摘要翻译： 提供了包括两个元件或组件的套件或系统。 第一组分（i）是包含核酸序列和至少一个原型间隔子的选择性组分。 第二组分（ii）包括至少一种包含至少一个cas基因和至少一个CRISPR阵列的敏化组分。 CRISPR的至少一个间隔区靶向包含在细菌的致病基因内的原型间隔区，以特异性灭活所述细菌中的所述致病基因，并且其中所述CRISPR的至少一个间隔区靶向包含在所述选择性组分中的原型间隔区 （i）的化合物，以特异性使所述选择性组分失活。 还提供了使用本发明的组分或试剂盒干扰细菌之间致病基因的水平转移并用于预防由细菌感染引起的哺乳动物受试者中的病理状况的方法。