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    METHODS FOR STRUCTURAL DETERMINATION OF SELENIUM DERIVATIZED NUCLEIC ACID COMPLEXES
    1.
    发明公开
    METHODS FOR STRUCTURAL DETERMINATION OF SELENIUM DERIVATIZED NUCLEIC ACID COMPLEXES 审中-公开
    VERFAHREN ZUR STRUKTURBESTIMMUNG VON SELENDERIVATISITENTENNUKLEINSÄUREKOMPLEXEN

    公开(公告)号：EP3071590A1

    公开(公告)日：2016-09-28

    申请号：EP14863946.1

    申请日：2014-11-21

    申请人： SeNA Research, Inc.

    发明人： HUANG, Zhen

    IPC分类号： C07K1/14 ,  C07K1/30 ,  G01N33/53 ,  G01N33/68

    CPC分类号： A61K47/4823 ,  A61K38/465 ,  A61K47/61 ,  C07K1/30 ,  C12N9/22 ,  C12N9/96 ,  C12Y301/26004 ,  G01N33/5308 ,  G06F19/00 ,  G16H50/50 ,  Y02A90/26

    摘要： Methods for crystallizing a molecule of interest, such as a polypeptide, in complex with nucleic acid, including contacting the molecule of interest with selenium-derivatized nucleic acid and crystallizing the molecule of interest/selenium-derivatized nucleic acid complex are provided. Methods for determining the X-ray crystal structure of molecule of interest/selenium-derivatized nucleic acid complexes are also provided. Typically, the method of X-ray crystal structural determination includes selenium single-wavelength anomalous phasing of the selenium-derivatized nucleic acid. In some embodiments the phases for the X-ray crystal structure of the molecule of interest are not provided from another crystal. Also disclosed are methods of affecting a biological process by administering a functional nucleic acid to a cell or a subject and/or by bringing into contact a nuclease and a functional nucleic acid, where the functional nucleic acid is selenium-derivatized nucleic acid.

    摘要翻译： 提供了与核酸复合的目标分子（例如多肽）结晶的方法，包括将感兴趣的分子与硒衍生的核酸接触并结晶感兴趣的分子/硒衍生的核酸复合物。 还提供了用于确定目的分子的X射线晶体结构/硒衍生的核酸复合物的方法。 通常，X射线晶体结构测定的方法包括硒衍生化核酸的硒单波长异常定相。 在一些实施方案中，感兴趣的分子的X射线晶体结构的相不是从另一晶体提供的。 还公开了通过向细胞或受试者施用功能性核酸和/或通过使功能性核酸为硒衍生化的核酸与核酸酶和功能性核酸接触来影响生物学过程的方法。

    Sequence-specific engineered ribonuclease H and the method for determining the sequence preference of DNA-RNA hybrid binding proteins
    2.
    发明公开
    Sequence-specific engineered ribonuclease H and the method for determining the sequence preference of DNA-RNA hybrid binding proteins 审中-公开
    序列特异性修饰核糖核酸酶H和方法，用于确定DNA-RNA杂交序列优先结合蛋白质的

    公开(公告)号：EP2857506A3

    公开(公告)日：2015-04-29

    申请号：EP14180902.0

    申请日：2012-06-07

    申请人： Miedzynarodowy Instytut Biologii Molekularnej I Komorkowej

    发明人： Bujnicki, Janusz Marek ,  Sulej, Agata Agnieszka ,  Skowronek, Krzysztof Jerzy ,  Nowotny, Marcin

    IPC分类号： C12N9/22

    CPC分类号： C12N9/22 ,  C07K14/435 ,  C12Y301/26004

    摘要： The present invention relates to the of determining the sequence preference of DNA-RNA hybrid binding protein or its domain which comprises the following steps:
    a) contacting of the purified protein or its domain, with a mixture of a library of DNA-RNA hybrids substrates, wherein DNA-RNA hybrid substrates comprise randomized sequences in the central part, preferably randomized in 9 or 10 nucleotide positions, with flanking sequences fixed and allowing a tested protein or its domain to bind the sequence to which it has an affinity;
    b) separating unbound DNA-RNA hybrids by immobilization of protein or its domain, with bound DNA-RNA hybrid, to the resin, preferably glutathione agarose;
    c) removing the unbound hybrids;
    d) isolating the recombinant protein, together with the associated DNA-RNA hybrids,
    e) amplification of the isolated hybrid using PCR, preferably RT-PCR, wherein in the amplification reaction the primers complementary to invariant sequences flanking the randomized region are used, and wherein during the PCR reaction on one of the primer the sequence of RNA polymerase promoter is added in order to obtain double-stranded DNA for in vitro transcription with RNA polymerase;
    f) reverse transcription using reverse transcriptase that does not possess the RNase H activity and a DNA primer complementary to the 3' end of the RNA template in order to obtain a DNA-RNA hybrid..

    NEW COMPACT SCAFFOLD OF CAS9 IN THE TYPE II CRISPR SYSTEM
    3.
    发明公开
    NEW COMPACT SCAFFOLD OF CAS9 IN THE TYPE II CRISPR SYSTEM 审中-公开
    关闭KOMPAKTESGERÜSTVON CAS9 IM TYP-II-CRISPR系统

    公开(公告)号：EP3004339A2

    公开(公告)日：2016-04-13

    申请号：EP14730473.7

    申请日：2014-05-28

    申请人： Cellectis

    发明人： DUCHATEAU, Philippe ,  BERTONATI, Claudia

    IPC分类号： C12N9/22 ,  C12N15/63 ,  C12N5/0783

    CPC分类号： C12N15/907 ,  C12N9/22 ,  C12N15/1082 ,  C12N15/63 ,  C12N15/902 ,  C12Y301/00 ,  C12Y301/26004

    摘要： The present invention is in the field of CRISPR-Cas system for genome targeting. The present invention relates to new engineered Cas9 scaffolds and uses thereof. More particularly, the present invention relates to methods for genome targeting, cell engineering and therapeutic application. The present invention also relates to vectors, compositions and kits in which the new Cas9 scaffolds of the present invention are used.

    摘要翻译： 本发明涉及用于基因组靶向的CRISPR-Cas系统。 本发明涉及新的工程化Cas9支架及其用途。 更具体地，本发明涉及用于基因组靶向，细胞工程和治疗应用的方法。 本发明还涉及使用本发明的新的Cas9支架的载体，组合物和试剂盒。

    METHODS FOR STRUCTURAL DETERMINATION OF SELENIUM DERIVATIZED NUCLEIC ACID COMPLEXES
    8.
    发明公开
    METHODS FOR STRUCTURAL DETERMINATION OF SELENIUM DERIVATIZED NUCLEIC ACID COMPLEXES 审中-公开
    VERFAHREN ZUR STRUKTURBESTIMMUNG VON SELENDERIVATISIERTENNUKLEINSÄARDKOMPLEXEN

    公开(公告)号：EP3071590A4

    公开(公告)日：2017-07-19

    申请号：EP14863946

    申请日：2014-11-21

    申请人： SENA RES INC

    发明人： HUANG ZHEN

    IPC分类号： A61K38/46 ,  C07K1/14 ,  C07K1/30 ,  C12N9/22 ,  C12N9/96 ,  G01N33/53 ,  G01N33/68 ,  G06F19/00

    CPC分类号： A61K47/4823 ,  A61K38/465 ,  A61K47/61 ,  C07K1/30 ,  C12N9/22 ,  C12N9/96 ,  C12Y301/26004 ,  G01N33/5308 ,  G06F19/3437 ,  G16H50/50 ,  Y02A90/26

    摘要： Methods for crystallizing a molecule of interest, such as a polypeptide, in complex with nucleic acid, including contacting the molecule of interest with selenium-derivatized nucleic acid and crystallizing the molecule of interest/selenium-derivatized nucleic acid complex are provided. Methods for determining the X-ray crystal structure of molecule of interest/selenium-derivatized nucleic acid complexes are also provided. Typically, the method of X-ray crystal structural determination includes selenium single-wavelength anomalous phasing of the selenium-derivatized nucleic acid. In some embodiments the phases for the X-ray crystal structure of the molecule of interest are not provided from another crystal. Also disclosed are methods of affecting a biological process by administering a functional nucleic acid to a cell or a subject and/or by bringing into contact a nuclease and a functional nucleic acid, where the functional nucleic acid is selenium-derivatized nucleic acid.

    摘要翻译： 提供了使与核酸复合的感兴趣的分子例如多肽结晶的方法，包括使感兴趣的分子与硒衍生化的核酸接触并结晶目的分子/硒衍生的核酸复合物。 还提供了确定感兴趣的分子/硒衍生的核酸复合物的X射线晶体结构的方法。 通常，X射线晶体结构测定的方法包括硒衍生化的核酸的硒单波长异相定位。 在一些实施方案中，感兴趣分子的X射线晶体结构的相不是从另一种晶体提供的。 还公开了通过将功能性核酸给予细胞或受试者和/或通过使核酸酶和功能性核酸接触来影响生物过程的方法，其中功能性核酸是硒衍生的核酸。

    Sequence-specific engineered ribonuclease H and the method for determining the sequence preference of DNA-RNA hybrid binding proteins
    10.
    发明公开
    Sequence-specific engineered ribonuclease H and the method for determining the sequence preference of DNA-RNA hybrid binding proteins 审中-公开
    序列特异性修饰核糖核酸酶H和方法，用于确定DNA-RNA杂交序列优先结合蛋白质的

    公开(公告)号：EP2857506A2

    公开(公告)日：2015-04-08

    申请号：EP14180902.0

    申请日：2012-06-07

    申请人： Miedzynarodowy Instytut Biologii Molekularnej I Komorkowej

    发明人： Bujnicki, Janusz Marek ,  Sulej, Agata Agnieszka ,  Skowronek, Krzysztof Jerzy ,  Nowotny, Marcin

    IPC分类号： C12N9/22

    CPC分类号： C12N9/22 ,  C07K14/435 ,  C12Y301/26004

    摘要： The present invention relates to the of determining the sequence preference of DNA-RNA hybrid binding protein or its domain which comprises the following steps:
    a) contacting of the purified protein or its domain, with a mixture of a library of DNA-RNA hybrids substrates, wherein DNA-RNA hybrid substrates comprise randomized sequences in the central part, preferably randomized in 9 or 10 nucleotide positions, with flanking sequences fixed and allowing a tested protein or its domain to bind the sequence to which it has an affinity;
    b) separating unbound DNA-RNA hybrids by immobilization of protein or its domain, with bound DNA-RNA hybrid, to the resin, preferably glutathione agarose;
    c) removing the unbound hybrids;
    d) isolating the recombinant protein, together with the associated DNA-RNA hybrids,
    e) amplification of the isolated hybrid using PCR, preferably RT-PCR, wherein in the amplification reaction the primers complementary to invariant sequences flanking the randomized region are used, and wherein during the PCR reaction on one of the primer the sequence of RNA polymerase promoter is added in order to obtain double-stranded DNA for in vitro transcription with RNA polymerase;
    f) reverse transcription using reverse transcriptase that does not possess the RNase H activity and a DNA primer complementary to the 3' end of the RNA template in order to obtain a DNA-RNA hybrid..

    摘要翻译： 本发明涉及DNA-RNA杂交物结合蛋白或其结构域的确定性挖掘的序列偏好其包括以下步骤：a）使纯化的蛋白质或其结构域，具有DNA-RNA杂交基板的文库的混合物 ，worin DNA-RNA杂交基板包含随机序列在中心部分，优选地随机分为9个或10个核苷酸位置，与侧翼序列和固定的允许一个测试蛋白质或其结构域的序列结合到由它来亲和力; b）在蛋白质或其结构域的固定化分离未结合的DNA-RNA杂交体，具有结合的DNA-RNA杂交，以该树脂，优选谷胱甘肽琼脂糖; c）除去未结合的杂种; d）在使用PCR，优选RT-PCR，worin在扩增反应中的引物与侧翼于随机化区域不变序列互补的分离的杂交体的相关的DNA-RNA杂交体，E）扩增分离重组蛋白质，一起被使用，并且 上的RNA聚合酶启动子的序列，以获得用于在体外与RNA聚合酶转录的双链DNA中加入引物中的一个PCR反应期间worin; F）使用逆转录酶没有不具有RNA酶H活性和DNA引物，以便获得一个DNA-RNA杂交到模板RNA的3“末端互补的反转录..