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    Mutagenesis method using polyethylene glycol mediated introduction of mutagenic nucleobases into plant protoplasts
    4.
    发明授权
    Mutagenesis method using polyethylene glycol mediated introduction of mutagenic nucleobases into plant protoplasts 有权
    诱变方法使用聚乙二醇介导诱变核碱基植入植物原生质体

    公开(公告)号：US09365860B2

    公开(公告)日：2016-06-14

    申请号：US13551143

    申请日：2012-07-17

    申请人： Paul Bundock ,  Michiel Theodoor Jan De Both ,  Franck Lhuissier

    发明人： Paul Bundock ,  Michiel Theodoor Jan De Both ,  Franck Lhuissier

    IPC分类号： C12N15/82

    CPC分类号： C12N15/8206 ,  C12N15/8213

    摘要： Method for targeted alteration of a duplex acceptor DNA sequence in a plant cell protoplast, comprising combining the duplex acceptor DNA sequence with a donor mutagenic nucleobase, wherein the duplex acceptor DNA sequence contains a first DNA sequence and a second DNA sequence which is the complement of the first DNA sequence and wherein the donor mutagenic nucleobase comprises at least one mismatch with respect to the duplex acceptor DNA sequence to be altered, preferably with respect to the first DNA sequence, wherein the method further comprises a step of introducing the donor mutagenic nucleobase into the cell protoplasts using polyethylene glycol (PEG) mediated transformation and the use of PEG protoplast transformation for enhancing the rate of targeted mutagenesis.

    摘要翻译： 用于靶向改变植物细胞原生质体中双链体受体DNA序列的方法，包括将双链体受体DNA序列与供体诱变核碱基组合，其中双链体受体DNA序列含有第一DNA序列和第二DNA序列， 所述第一DNA序列，并且其中所述供体诱变核碱基相对于待改变的双链体受体DNA序列至少包含一个错配，优选相对于所述第一DNA序列，其中所述方法还包括将供体诱变核碱基引入 使用聚乙二醇（PEG）介导的转化的细胞原生质体和使用PEG原生质体转化来提高定向诱变的速率。

    TARGETED ALTERATION OF DNA WITH OLIGONUCLEOTIDES
    5.
    发明申请
    TARGETED ALTERATION OF DNA WITH OLIGONUCLEOTIDES 有权
    DNA与寡核苷酸的靶向修饰

    公开(公告)号：US20130330774A1

    公开(公告)日：2013-12-12

    申请号：US13991065

    申请日：2011-11-25

    申请人： Michiel Theodoor Jan De Both ,  Tomoyuki Furukawa

    发明人： Michiel Theodoor Jan De Both ,  Tomoyuki Furukawa

    IPC分类号： C12N15/10

    CPC分类号： C12N15/1024 ,  C12N15/102 ,  C12N15/1082

    摘要： The current invention relates to a method for targeted alteration of acceptor DNA, for example duplex acceptor DNA. The method comprises use of at least two oligonucleotides, each oligonucleotide having at least one mismatch relative to the targeted (duplex) acceptor DNA. The mismatch of the first oligonucleotide is directed to a nucleotide at a position in the first strand of the duplex and the mismatch of the second oligonucleotide is directed to the nucleotide in the second strand that occupies the complementary position in the duplex acceptor DNA (e.g. forms a base-pair with the nucleotide in the first strand). These mismatches are located at specific positions within said oligonucleotides. Also provided is a kit that comprises instructions for performing the method according to the inventions, and in a preferred embodiment, comprises oligonucleotides suitable for use in the method.

    摘要翻译： 本发明涉及用于靶向改变受体DNA的方法，例如双链受体DNA。 该方法包括使用至少两种寡核苷酸，每种寡核苷酸相对于靶向（双链体）受体DNA具有至少一个失配。 第一寡核苷酸的不匹配指向双链体第一链中位置的核苷酸，第二寡核苷酸的错配指向第二链中位于双链受体DNA中的互补位置的核苷酸（例如形式 与第一链中的核苷酸的碱基对）。 这些错配位于所述寡核苷酸内的特定位置。 还提供了包括用于执行根据本发明的方法的指令的试剂盒，并且在优选的实施方案中，包含适合用于该方法的寡核苷酸。

    MUTAGENESIS METHOD USING POLYETHYLENE GLYCOL MEDIATED INTRODUCTION OF MUTAGENIC NUCLEOBASES INTO PLANT PROTOPLASTS
    6.
    发明申请
    MUTAGENESIS METHOD USING POLYETHYLENE GLYCOL MEDIATED INTRODUCTION OF MUTAGENIC NUCLEOBASES INTO PLANT PROTOPLASTS 有权
    使用聚乙二醇将诱导性核酸引入植物原核生物的诱变方法

    公开(公告)号：US20120282699A1

    公开(公告)日：2012-11-08

    申请号：US13551143

    申请日：2012-07-17

    申请人： Paul BUNDOCK ,  Michiel Theodoor Jan De Both ,  Franck Lhuissier

    发明人： Paul BUNDOCK ,  Michiel Theodoor Jan De Both ,  Franck Lhuissier

    IPC分类号： C12N15/82

    CPC分类号： C12N15/8206 ,  C12N15/8213

    摘要： Method for targeted alteration of a duplex acceptor DNA sequence in a plant cell protoplast, comprising combining the duplex acceptor DNA sequence with a donor mutagenic nucleobase, wherein the duplex acceptor DNA sequence contains a first DNA sequence and a second DNA sequence which is the complement of the first DNA sequence and wherein the donor mutagenic nucleobase comprises at least one mismatch with respect to the duplex acceptor DNA sequence to be altered, preferably with respect to the first DNA sequence, wherein the method further comprises a step of introducing the donor mutagenic nucleobase into the cell protoplasts using polyethylene glycol (PEG) mediated transformation and the use of PEG protoplast transformation for enhancing the rate of targeted mutagenesis.

    摘要翻译： 用于靶向改变植物细胞原生质体中双链体受体DNA序列的方法，包括将双链体受体DNA序列与供体诱变核碱基组合，其中双链体受体DNA序列含有第一DNA序列和第二DNA序列， 所述第一DNA序列，并且其中所述供体诱变核碱基相对于待改变的双链体受体DNA序列至少包含一个错配，优选相对于所述第一DNA序列，其中所述方法还包括将供体诱变核碱基引入 使用聚乙二醇（PEG）介导的转化的细胞原生质体和使用PEG原生质体转化来提高定向诱变的速率。

    MUTAGENESIS METHOD USING POLYETHYLENE GLYCOL MEDIATED INTRODUCTION OF MUTAGENIC NUCLEOBASES INTO PLANT PROTOPLASTS
    7.
    发明申请
    MUTAGENESIS METHOD USING POLYETHYLENE GLYCOL MEDIATED INTRODUCTION OF MUTAGENIC NUCLEOBASES INTO PLANT PROTOPLASTS 审中-公开
    使用聚乙二醇将诱导性核酸引入植物原核生物的诱变方法

    公开(公告)号：US20100291684A1

    公开(公告)日：2010-11-18

    申请号：US12809384

    申请日：2007-12-21

    申请人： Paul Bundock ,  Michiel Theodoor Jan De Both ,  Franck Lhuissier

    发明人： Paul Bundock ,  Michiel Theodoor Jan De Both ,  Franck Lhuissier

    IPC分类号： C12N15/82

    CPC分类号： C12N15/8206 ,  C12N15/8213

    摘要： Method for targeted alteration of a duplex acceptor DNA sequence in a plant cell protoplast, comprising combining the duplex acceptor DNA sequence with a donor mutagenic nucleobase, wherein the duplex acceptor DNA sequence contains a first DNA sequence and a second DNA sequence which is the complement of the first DNA sequence and wherein the donor mutagenic nucleobase comprises at least one mismatch with respect to the duplex acceptor DNA sequence to be altered, preferably with respect to the first DNA sequence, wherein the method further comprises a step of introducing the donor mutagenic nucleobase into the cell protoplasts using polyethylene glycol (PEG) mediated transformation and the use of PEG protoplast transformation for enhancing the rate of targeted mutagenesis.

    摘要翻译： 用于靶向改变植物细胞原生质体中的双链体受体DNA序列的方法，包括将双链体受体DNA序列与供体诱变核碱基组合，其中双链体受体DNA序列含有第一DNA序列和第二DNA序列， 所述第一DNA序列，并且其中所述供体诱变核碱基相对于待改变的双链体受体DNA序列至少包含一个错配，优选相对于所述第一DNA序列，其中所述方法还包括将供体诱变核碱基引入 使用聚乙二醇（PEG）介导的转化的细胞原生质体和使用PEG原生质体转化来提高定向诱变的速率。

    Targeted alteration of DNA with oligonucleotides
    10.
    发明授权
    Targeted alteration of DNA with oligonucleotides 有权
    用寡核苷酸靶向改变DNA

    公开(公告)号：US09518258B2

    公开(公告)日：2016-12-13

    申请号：US13991065

    申请日：2011-11-25

    申请人： Michiel Theodoor Jan De Both ,  Tomoyuki Furukawa

    发明人： Michiel Theodoor Jan De Both ,  Tomoyuki Furukawa

    IPC分类号： C12N15/00 ,  C12N15/10

    CPC分类号： C12N15/1024 ,  C12N15/102 ,  C12N15/1082

    摘要： The current invention relates to a method for targeted alteration of acceptor DNA, for example duplex acceptor DNA. The method comprises use of at least two oligonucleotides, each oligonucleotide having at least one mismatch relative to the targeted (duplex) acceptor DNA. The mismatch of the first oligonucleotide is directed to a nucleotide at a position in the first strand of the duplex and the mismatch of the second oligonucleotide is directed to the nucleotide in the second strand that occupies the complementary position in the duplex acceptor DNA (e.g. forms a base-pair with the nucleotide in the first strand). These mismatches are located at specific positions within said oligonucleotides. Also provided is a kit that comprises instructions for performing the method according to the inventions, and in a preferred embodiment, comprises oligonucleotides suitable for use in the method.

    摘要翻译： 本发明涉及用于靶向改变受体DNA的方法，例如双链受体DNA。 该方法包括使用至少两种寡核苷酸，每种寡核苷酸相对于靶向（双链体）受体DNA具有至少一个失配。 第一寡核苷酸的不匹配指向双链体第一链中位置的核苷酸，第二寡核苷酸的错配指向第二链中位于双链受体DNA中的互补位置的核苷酸（例如形式 与第一链中的核苷酸的碱基对）。 这些错配位于所述寡核苷酸内的特定位置。 还提供了包括用于执行根据本发明的方法的指令的试剂盒，并且在优选的实施方案中，包含适合用于该方法的寡核苷酸。