公开(公告)号：WO2016137774A1

公开(公告)日：2016-09-01

申请号：PCT/US2016/017937

申请日：2016-02-15

Applicant: PIONEER HI-BRED INTERNATIONAL INC

Inventor： CIGAN, Andrew Mark ,  SVITASHEV, Sergei

IPC: C12N15/82 ,  C12N9/22 ,  A01H1/04

CPC classification number: C12N15/8237 ,  C12N9/22 ,  C12N15/8212 ,  C12N15/8213

Abstract: Compositions and methods are provided for regulated expression of a guide RNA/Cas endonuclease system in a plant cell, plant and seed. Compositions and methods are also provided for genome modification of a target sequence in the genome of a plant or plant cell. The methods and compositions employ a regulated 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.

Abstract translation: 提供组合物和方法用于在植物细胞，植物和种子中调控RNA / Cas内切核酸酶系统的表达。 还提供组合物和方法用于基因组修饰植物或植物细胞基因组中的靶序列。 所述方法和组合物使用调控的引导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。

公开(公告)号：WO2021016098A1

公开(公告)日：2021-01-28

申请号：PCT/US2020/042579

申请日：2020-07-17

Applicant: PIONEER HI-BRED INTERNATIONAL, INC.

Inventor： DAS, Amit ,  KANUGANTI, Satish ,  MOHANTY, Amitabh ,  SVITASHEV, Sergei ,  TAVVA, Venkata S

IPC: C12N9/22 ,  C12N15/10 ,  C12N15/82

Abstract: Methods and compositions are provided for the improvement of homology-directed repair of a double strand break in a plant cell, using concatemers of heterologous polynucleotides that are flanked by sequences capable of sequence hybridization with a guide RNA. In some aspects, the double strand break is created by an RNA-guided Cas endonuclease. The homology-directed repair of the double-strand break may include incorporation of a heterologous polynucleotide, for example a gene encoding a trait of agronomic importance. The homology-directed repair of the double-strand break may occur as a result of template-directed repair using a heterologous polynucleotide as a repair template.

公开(公告)号：WO2019217354A1

公开(公告)日：2019-11-14

申请号：PCT/US2019/031017

申请日：2019-05-07

Applicant: PIONEER HI-BRED INTERNATIONAL, INC.

Inventor： ANAND, Ajith ,  GORDON-KAMM, William James ,  KUMAR, Sandeep ,  LIU, Zhan-Bin ,  SVITASHEV, Sergei

IPC: A61K48/00 ,  C12N5/10 ,  C12N9/22 ,  C12N15/09 ,  C12N15/11 ,  C12N15/82

Abstract: Methods and compositions are provided for the improvement of homology-directed repair of a double strand break in a plant cell, via the use of a polynucleotide comprising sequences homologous to the target site. In some aspects, the double strand break is created by an RNA-guided Cas endonuclease. The homology-directed repair of the double-strand break may include incorporation of a heterologous polynucleotide, for example a gene encoding a trait of agronomic importance. The homology-directed repair of the double-strand break may occur as a result of template-directed repair using a polynucleotide repair template.

公开(公告)号：WO2015026886A1

公开(公告)日：2015-02-26

申请号：PCT/US2014/051781

申请日：2014-08-20

Applicant: E. I. DU PONT DE NEMOURS AND COMPANY ,  PIONEER HI-BRED INTERNATIONAL, INC.

Inventor： CIGAN, Andrew, Mark ,  FALCO, Saverio, Carl ,  LASSNER, Michael ,  LIU, Zhan-Bin ,  SVITASHEV, Sergei

IPC: C12N15/82

CPC classification number: 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

Abstract: Compositions and methods are provided for genome modification of a target sequence in the genome of a plant or plant cell, without incorporating a selectable transgene marker. 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 plant, plant cell or seed, without incorporating a selectable transgene marker. 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 guide polynucleotide / Cas endonuclease system are also disclosed. Compositions and methods are also provided for editing a nucleotide sequence in the genome of a cell, without incorporating a selectable transgene marker.

Abstract translation: 提供组合物和方法用于在植物或植物细胞的基因组中靶基因组的基因组修饰，而不加入可选择的转基因标记。 方法和组合物采用引导多核苷酸/ 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，而不需要并入可选择的转基因标记。 一旦识别出基因组靶位点，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。 还公开了利用引导多核苷酸/ 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，而不包含可选择的转基因标记。

公开(公告)号：WO2012129373A2

公开(公告)日：2012-09-27

申请号：PCT/US2012/030061

申请日：2012-03-22

Applicant: PIONEER HI-BRED INTERNATIONAL, INC. ,  E.I. DU PONT DE NEMOURS AND COMPANY ,  LASSNER, Michael ,  JANTZ, Derek ,  SMITH, James, Jefferson ,  CIGAN, Mark ,  FALCO, Carl ,  GAO, Huirong ,  LI, Zhongsen ,  LIU, Zhan-bin ,  SVITASHEV, Sergei

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

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

CPC classification number: C12N15/8202 ,  A01H1/06 ,  C12N9/22 ,  C12N15/8213

Abstract: 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.

Abstract translation: 公开了在植物中生产复合转基因性状基因座的方法，所述基因座包含在目的基因组区域中的至少两个改变的靶序列。 所述方法涉及使用两种或更多种双链断裂诱导剂，其中每一种可导致目的基因组区域中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。

公开(公告)号：WO2009154639A1

公开(公告)日：2009-12-23

申请号：PCT/US2008/071429

申请日：2008-07-29

Applicant: PIONEER HI-BRED INTERNATIONAL, INC. ,  ANANIEV, Evgueni ,  CHAMBERLIN, Mark, A. ,  GORDON-KAMM, William, J. ,  SVITASHEV, Sergei ,  WU, Chengcang

Inventor： ANANIEV, Evgueni ,  CHAMBERLIN, Mark, A. ,  GORDON-KAMM, William, J. ,  SVITASHEV, Sergei ,  WU, Chengcang

IPC: C12N15/82

CPC classification number: C12N15/8201 ,  C12N15/82

Abstract: Artificial plant minichromosomes comprising a functional centromere which specifically bind centromeric protein C (CENPC) and methods for making such minichromosomes are described.

Abstract translation: 描述了包含特异性结合着丝粒蛋白C（CENPC）的功能着丝粒的人造植物微染色体和制备这种微染色体的方法。

公开(公告)号：WO2007137114A2

公开(公告)日：2007-11-29

申请号：PCT/US2007/069139

申请日：2007-05-17

Applicant: PIONEER HI-BRED INTERNATIONAL, INC. ,  ANANIEV, Evgueni ,  CHAMBERLIN, Mark A. ,  GORDON-KAMM, William J. ,  SVITASHEV, Sergei ,  WU, Chengchang

Inventor： ANANIEV, Evgueni ,  CHAMBERLIN, Mark A. ,  GORDON-KAMM, William J. ,  SVITASHEV, Sergei ,  WU, Chengchang

IPC: C12N15/82

CPC classification number: C12N15/82 ,  C12N15/8201

Abstract: Artificial plant minichromosomes comprising a functional centromere which specifically bind centromeric protein C (CENPC) and methods for making such minichromosomes are described.

Abstract translation: 描述了包含特异性结合着丝粒蛋白C（CENPC）的功能着丝粒的人造植物微染色体和制备这种微染色体的方法。

公开(公告)号：WO2021141890A1

公开(公告)日：2021-07-15

申请号：PCT/US2021/012173

申请日：2021-01-05

Applicant: PIONEER HI-BRED INTERNATIONAL, INC.

Inventor： GAO, Huirong ,  SVITASHEV, Sergei

IPC: A61K38/46 ,  C12N9/22 ,  C12N15/11 ,  C12N15/113 ,  C12N15/29 ,  C12N15/90

Abstract: Compositions and methods are provided for the excision and replacement of an endogenous polynucleotide, such as a gene, using CRISPR-Cas systems. In some aspects, the gene is flanked by specific nucleotides that are targets of homology-directed repair, for the insertion of a replacement polynucleotide. Also provided are methods and compositions for replacement of polynucleotides in genomes comprising highly repetitive regions, and for improving the phenotype of an organism.

公开(公告)号：WO2017070032A1

公开(公告)日：2017-04-27

申请号：PCT/US2016/057279

申请日：2016-10-17

Applicant: PIONEER HI-BRED INTERNATIONAL, INC.

Inventor： CIGAN, Andrew Mark ,  SVITASHEV, Sergei

IPC: C12N9/22 ,  C12N15/01 ,  C12N15/82 ,  C12N15/90

CPC classification number: C12N15/8206 ,  C07K2319/09 ,  C12N9/22 ,  C12N15/8207 ,  C12N15/8241 ,  C12N15/902

Abstract: Compositions and methods are provided for restoring function to a non-functional gene product in the genome of a cell. The methods and compositions employ a guide polynucleotide /Cas endonuclease system to restore function to a non-functional gene product and to provide an effective system for modifying or altering target sites within the genome of a plant, plant cell or seed. The present disclosure also describes methods for modifying a nucleotide sequence in the genome of a cell using a restored functional selectable marker, as well as methods for editing a nucleotide sequence in the genome a cell without introducing a polynucleotide modification template into said cell. Compositions and methods are also provided for DNA free delivery of Cas endonucleases, sgRNAs and guide RNA/Cas complexes.

Abstract translation: 提供组合物和方法用于恢复细胞基因组中的非功能性基因产物的功能。 所述方法和组合物使用引导性多核苷酸/ Cas核酸内切酶系统来恢复非功能性基因产物的功能，并提供用于修饰或改变植物，植物细胞或种子的基因组内的靶位点的有效系统。 本公开还描述了使用修复的功能选择标记修饰细胞基因组中的核苷酸序列的方法，以及编码细胞基因组中核苷酸序列而不将多核苷酸修饰模板引入所述细胞的方法。 还提供了用于Cas内切核酸酶，sgRNA和指导RNA / Cas复合物的DNA免费递送的组合物和方法。

公开(公告)号：WO2015026883A1

公开(公告)日：2015-02-26

申请号：PCT/US2014/051778

申请日：2014-08-20

Applicant: E. I. DU PONT DE NEMOURS AND COMPANY ,  PIONEER HI-BRED INTERNATIONAL, INC.

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

IPC: C12N15/82

CPC classification number: 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

Abstract: 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.

Abstract translation: 提供组合物和方法用于基因组修饰植物或植物细胞基因组中的靶序列。 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。 一旦识别出基因组靶位点，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。