公开(公告)号：EP3036334A1

公开(公告)日：2016-06-29

申请号：EP14761479.6

申请日：2014-08-20

申请人： E. I. du Pont de Nemours and Company ,  Pioneer Hi-Bred International, Inc.

发明人： CIGAN, Andrew Mark ,  FALCO, Saverio Carl ,  LI, Zhongsen ,  LIU, Zhan-Bin ,  YOUNG, Joshua K.

IPC分类号： C12N15/82 ,  C07K14/415

CPC分类号： C12N15/8216 ,  C12N15/113 ,  C12N15/81 ,  C12N15/8213 ,  C12N15/8241 ,  C12N15/8245 ,  C12N15/8247 ,  C12N15/8261 ,  C12N15/8273 ,  C12N15/8274 ,  C12N15/8279 ,  C12N15/8286 ,  C12N2310/10 ,  C12N2310/20 ,  C12N2310/315 ,  C12N2310/3231 ,  C12N2310/3341 ,  C12Q1/6895 ,  C12Q2600/13 ,  C12Q2600/156 ,  C12N2310/321 ,  C12N2310/3521 ,  C12N2310/322 ,  C12N2310/3533

摘要： Compositions and methods are provided for genome modification of a target sequence in the genome of a plant or plant cell. The methods and compositions employ a guide RNA/Cas endonuclease system to provide an effective system for modifying or altering target sites within the genome of a plant, plant cell or seed. Also provided are compositions and methods employing a guide polynucleotide/Cas endonuclease system for genome modification of a nucleotide sequence in the genome of a cell or organism, for gene editing, and/or for inserting or deleting a polynucleotide of interest into or from the genome of a cell or organism. Once a genomic target site is identified, a variety of methods can be employed to further modify the target sites such that they contain a variety of polynucleotides of interest. Breeding methods and methods for selecting plants utilizing a two component RNA guide and Cas endonuclease system are also disclosed. Compositions and methods are also provided for editing a nucleotide sequence in the genome of a cell.

摘要翻译： 提供组合物和方法用于植物或植物细胞基因组中靶序列的基因组修饰。 方法和组合物采用指导性RNA / Cas内切核酸酶系统来提供用于修饰或改变植物，植物细胞或种子基因组内的靶位点的有效系统。 还提供了使用引导多核苷酸/ Cas内切核酸酶系统用于细胞或生物体基因组中的核苷酸序列进行基因组修饰，用于基因编辑和/或用于在基因组中插入或缺失目标多核苷酸或从基因组中插入或缺失目的多核苷酸的组合物和方法 的细胞或生物体。 一旦确定了基因组靶位点，就可以使用多种方法来进一步修饰靶位点，使得它们含有多种感兴趣的多核苷酸。 还公开了利用双组分RNA引物和Cas内切核酸酶系统选择植物的育种方法和方法。 还提供组合物和方法用于编辑细胞基因组中的核苷酸序列。

公开(公告)号：EP3036332B1

公开(公告)日：2020-06-24

申请号：EP14761477.0

申请日：2014-08-20

申请人： E. I. du Pont de Nemours and Company ,  Pioneer Hi-Bred International, Inc.

发明人： CIGAN, Andrew ,  FALCO, Saverio Carl ,  GAO, Huirong ,  LI, Zhongsen ,  LIU, Zhan-Bin ,  LYZNIK, L. Aleksander ,  SHI, Jinrui ,  SVITASHEV, Sergei ,  YOUNG, Joshua K.

IPC分类号： C12N15/82

公开(公告)号：EP3036332A1

公开(公告)日：2016-06-29

申请号：EP14761477.0

申请日：2014-08-20

申请人： E. I. du Pont de Nemours and Company ,  Pioneer Hi-Bred International, Inc.

发明人： CIGAN, Andrew ,  FALCO, Saverio Carl ,  GAO, Huirong ,  LI, Zhongsen ,  LIU, Zhan-Bin ,  LYZNIK, L. Aleksander ,  SHI, Jinrui ,  SVITASHEV, Sergei ,  YOUNG, Joshua K.

IPC分类号： C12N15/82

摘要： Compositions and methods are provided for genome modification of a target sequence in the genome of a cell. The methods and compositions employ a guide polynucleotide/Cas endonuclease system to provide an effective system for modifying or altering target sites within the genome of a cell or organism. Once a genomic target site is identified, a variety of methods can be employed to further modify the target sites such that they contain a variety of polynucleotides of interest. Compositions and methods are also provided for editing a nucleotide sequence in the genome of a cell. Breeding methods and methods for selecting plants utilizing a two component RNA polynucleotide and Cas endonuclease system are also disclosed.

摘要翻译： 提供组合物和方法用于植物或植物细胞基因组中靶序列的基因组修饰。 方法和组合物采用指导性RNA / Cas内切核酸酶系统来提供用于修饰或改变植物，植物细胞或种子基因组内的靶位点的有效系统。 还提供了使用引导多核苷酸/ Cas内切核酸酶系统用于细胞或生物体基因组中的核苷酸序列进行基因组修饰，用于基因编辑和/或用于在基因组中插入或缺失目标多核苷酸或从基因组中插入或缺失目的多核苷酸的组合物和方法 的细胞或生物体。 一旦确定了基因组靶位点，就可以使用多种方法来进一步修饰靶位点，使得它们含有多种感兴趣的多核苷酸。 还公开了利用双组分RNA引物和Cas内切核酸酶系统选择植物的育种方法和方法。 还提供组合物和方法用于编辑细胞基因组中的核苷酸序列。

公开(公告)号：EP3611268A1

公开(公告)日：2020-02-19

申请号：EP19199945.7

申请日：2014-08-20

申请人： E. I. du Pont de Nemours and Company ,  Pioneer Hi-Bred International, Inc.

发明人： CIGAN, Andrew ,  FALCO, Saverio Carl ,  GAO, Huirong ,  LI, Zhongsen ,  LIU, Zhan-Bin ,  LYZNIK, L. Aleksander ,  SHI, Jinrui ,  SVITASHEV, Sergei ,  YOUNG, Joshua K.

IPC分类号： C12N15/82 ,  C12N15/113

摘要： Compositions and methods are provided for genome modification of a target sequence in the genome of a plant or plant cell. The methods and compositions employ a guide RNA/Cas endonuclease system to provide an effective system for modifying or altering target sites within the genome of a plant, plant cell or seed. Also provided are compositions and methods employing a guide polynucleotide/Cas endonuclease system for genome modification of a nucleotide sequence in the genome of a cell or organism, for gene editing, and/or for inserting or deleting a polynucleotide of interest into or from the genome of a cell or organism. Once a genomic target site is identified, a variety of methods can be employed to further modify the target sites such that they contain a variety of polynucleotides of interest. Breeding methods and methods for selecting plants utilizing a two component RNA guide and Cas endonuclease system are also disclosed. Compositions and methods are also provided for editing a nucleotide sequence in the genome of a cell.

公开(公告)号：EP2643464B1

公开(公告)日：2018-12-26

申请号：EP10784933.3

申请日：2010-11-24

申请人： Pioneer Hi-Bred International, Inc. ,  E. I. du Pont de Nemours and Company

发明人： CHARNE, David, George ,  CHEN, Wenpin ,  KOSCIELNY, Chadwick, Bruce ,  PATEL, Jayantilal, Devabhai ,  THOONEN, Ferdinand, Gerard ,  TULSIERAM, Lomas ,  ZHANG, Yongping ,  LI, Zhongsen

IPC分类号： C12N15/82

CPC分类号： C12N15/8275 ,  C12N15/8274

摘要： Compositions and methods related to transgenic glyphosate tolerant Brassica plants are provided. Specifically, the present invention provides Brassica plants having a DP-073496-4 event which imparts tolerance to glyphosate. The Brassica plant harboring the DP-073496-4 event at the recited chromosomal location comprises genomic/transgene junctions within SEQ ID NO: 2 or with genomic/transgene junctions as set forth in SEQ ID NO: 12 and/or 13. The characterization of the genomic insertion site of the event provides for an enhanced breeding efficiency and enables the use of molecular markers to track the transgene insert in the breeding populations and progeny thereof. Various methods and compositions for the identification, detection, and use of the event are provided.

公开(公告)号：EP2807257A1

公开(公告)日：2014-12-03

申请号：EP13702566.4

申请日：2013-01-24

申请人： Pioneer-Hi-Bred International, Inc. ,  E. I. Du Pont de Nemours and Company

发明人： FALCO, Saverio Carl ,  LASSNER, Michael ,  LI, Zhongsen ,  SCELONGE, Christopher J.

IPC分类号： C12N15/82 ,  C12N9/22

CPC分类号： A01H5/10 ,  A01H1/02 ,  C12N9/22 ,  C12N15/8213

摘要： Compositions and methods are provided for stacking multiple independent transgenic loci into the genome of a plant. Compositions include plants, seeds or plant cells comprising at least one transgenic target site and at least one genomic locus of interest integrated at different genomic sites within a genomic window. Plant breeding techniques can be employed such that the transgenic target site and the genomic locus of interest can be bred together. In this way, multiple independent transgene integrations can be generated within a genomic window to create a complex trait locus. The complex trait locus is designed such that the transgenic target sites and/or genomic loci of interest can segregate independently of each other, thus providing the benefit of altering a complex trait locus by breeding-in and breeding-away specific elements. Various methods can also be employed to modify the target sites such that they contain a variety of polynucleotides of interest.

摘要翻译： 提供了将多个独立的转基因基因座堆叠到植物基因组中的组合物和方法。 组合物包括植物，种子或植物细胞，其包含至少一个转基因靶位点和在基因组窗口内的不同基因组位点处整合的至少一个感兴趣的基因组。 可以使用植物育种技术，使得转基因靶位点和感兴趣的基因组座位可以一起繁殖。 以这种方式，可以在基因组窗口内产生多个独立的转基因整合以产生复杂的性状基因座。 复杂特征位点被设计成使得目的基因座和/或基因座位点彼此独立地分离，从而提供通过繁殖和遗传特异性元件改变复杂性状基因座的益处。 还可以使用各种方法来修饰靶位点，使得它们含有多种感兴趣的多核苷酸。

公开(公告)号：EP2739739A1

公开(公告)日：2014-06-11

申请号：EP12741436.5

申请日：2012-07-18

申请人： E. I. Du Pont de Nemours and Company ,  Pioneer Hi-Bred International, Inc.

发明人： CIGAN, Andrew Mark ,  FALCO, Saverio Carl ,  GAO, Huirong ,  LASSNER, Michael W. ,  LI, Zhongsen ,  LIU, Zhan-Bin ,  LYZNIK, Leszek A. ,  PETERSON, David J. ,  SCELONGE, Christopher Jay

IPC分类号： C12N15/82 ,  A01H5/00

CPC分类号： C12N15/8213

摘要： Compositions and methods are provided for the targeted integration of a polynucleotide sequence of interest into the genome of a plant or plant cell. The methods and compositions employ recognition sites for endonucleases and endonucleases in combination with site-specific recombination sites/recombinases to provide an effective system for establishing target sites within the genome of a plant, plant cell or seed. Once such target sites are established, a variety of methods can be employed to further modify the target sites such that they contain a variety of polynucleotides of interest.

公开(公告)号：EP3274460A1

公开(公告)日：2018-01-31

申请号：EP16715935.9

申请日：2016-03-21

申请人： E. I. du Pont de Nemours and Company

发明人： LI, Zhongsen

IPC分类号： C12N15/82

CPC分类号： C12N15/8216 ,  C12N15/8213

摘要： The invention relates to gene expression regulatory sequences from soybean, specifically to promoters of U6 small nuclear RNA genes, and fragments thereof, and their use in promoting the expression of one or more heterologous nucleic acid fragments in a constitutive manner in plants. The invention further discloses compositions, polynucleotide constructs, transformed host cells, transgenic plants and seeds containing the recombinant construct with the promoter, and methods for preparing and using the same.

公开(公告)号：EP2373798A1

公开(公告)日：2011-10-12

申请号：EP09775058.2

申请日：2009-12-16

申请人： E. I. du Pont de Nemours and Company

发明人： FALCO, Saverio, Carl ,  LI, Zhongsen

IPC分类号： C12N15/82 ,  A01H5/00 ,  A01H5/10

CPC分类号： C12N15/8213

摘要： A targeting method is described that allows precise cassette replacement at a previously characterized genetic locus. A target DNA construct containing a pair of incompatible
FRT sites flanking a target cassette was introduced into soybean by regular biolistic transformation. Transgenic events containing a single complete copy of the target site were then selected and retransformed with a donor DNA construct containing the identical pair of incompatible
FRT sites flanking a donor cassette. Precise DNA cassette exchange happened between the target cassette and the donor cassette via recombinase mediated cassette exchange (RMCE) so that the donor cassette was introduced at the exact genomic site previously occupied by the target cassette. Through repeated RMCE using additional incompatible
FRT sites, multiple groups of transgenes can be stacked at the same genomic locus.

公开(公告)号：EP2689019A2

公开(公告)日：2014-01-29

申请号：EP12712040.0

申请日：2012-03-22

申请人： Pioneer Hi-Bred International Inc. ,  E.I. Du Pont De Nemours and Company

发明人： LASSNER, Michael ,  JANTZ, Derek ,  SMITH, James, Jefferson ,  CIGAN, Mark ,  FALCO, Carl ,  GAO, Huirong ,  LI, Zhongsen ,  LIU, Zhan-bin ,  SVITASHEV, Sergei

IPC分类号： C12N15/82 ,  A01H1/06 ,  A01H5/10 ,  C12N15/55

CPC分类号： C12N15/8202 ,  A01H1/06 ,  C12N9/22 ,  C12N15/8213

摘要： Methods for producing in a plant a complex transgenic trait locus comprising at least two altered target sequences in a genomic region of interest are disclosed. The methods involve the use of two or more double-strand-break-inducing agents, each of which can cause a double-strand break in a target sequence in the genomic region of interest which results in an alteration in the target sequence. Also disclosed are complex transgenic trait loci in plants. A complex transgenic trait locus comprises at least two altered target sequences that are genetically linked to a polynucleotide of interest. Plants, plant cells, plant parts, and seeds comprising one or more complex transgenic trait loci are also disclosed.

摘要翻译： 公开了一种在植物中生产复合转基因性状基因座的方法，该基因座包含至少两个在目的基因组区域中改变的靶序列。 所述方法包括使用两种或更多种双链断裂诱导剂，其中每一种可引起目的基因组区域中的靶序列中的双链断裂，导致目标序列的改变。 还公开了植物中的复杂的转基因性状基因座。 复杂的转基因性状基因座包含与感兴趣的多核苷酸遗传连锁的至少两个改变的靶序列。 还公开了植物，植物细胞，植物部分和包含一个或多个复合转基因性状基因座的种子。