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1. 发明申请

WO2016040030A1 GENERATION OF SITE-SPECIFIC-INTEGRATION SITES FOR COMPLEX TRAIT LOCI IN CORN AND SOYBEAN, AND METHODS OF USE 审中-公开
标题翻译：玉米和大豆中复合物种位点特异性整合位点的生成及其使用方法
公开(公告)号：WO2016040030A1
公开(公告)日：2016-03-17
申请号：PCT/US2015/047706
申请日：2015-08-31
申请人： E. I. DU PONT DE NEMOURS AND COMPANY , PIONEER HI-BRED INTERNATIONAL, INC.
发明人： CIGAN, Andrew Mark , GAO, Huirong , LIU, Zhan-Bin , MUTTI, Jasdeep , PODLICH, Dean , SCELONGE, Christopher
IPC分类号： C12N15/82 , A01H5/10
CPC分类号： C12N15/8213 , C12N15/8201 , C12N15/8241
摘要： Compositions and methods are provided for introducing transgenic target sites for Site Specific Integration (SSI) and/or polynucleotides of interest into at least one double-strand break target site of a double-strand-break inducing agent in a genomic window of a plant genome. Also provided are methods and compositions for producing a complex trait locus in a genomic window of a plant comprising at least one transgenic target site for site specific integration integrated in at least double-strand-break target site. The double-strand-break target site can be, but is not limited to, a target site for a zinc finger endonuclease, an engineered endonuclease, a meganuclease, a TALENs and/or a Cas endonuclease. The genomic window of said plant can comprise at least one genomic locus of interest such as a trait cassette, a transgene, a mutated gene, a native gene, an edited gene or a site-specific integration (SSI) target site.
摘要翻译：提供组合物和方法用于将位点特异性整合（SSI）和/或目标多核苷酸的转基因靶位点引入植物基因组的基因组窗口中双链断裂诱导剂的至少一个双链断裂靶位点。还提供了用于在植物的基因组窗口中产生复合特征基因座的方法和组合物，其包含至少一个整合在至少双链断裂靶位点中的位点特异性整合的转基因靶位点。双链断裂靶位点可以是但不限于锌指内切核酸酶，工程内切核酸酶，大范围核酸酶，TALENs和/或Cas内切核酸酶的靶位点。所述植物的基因组窗可以包含至少一个感兴趣的基因座，例如性状盒，转基因，突变基因，天然基因，编辑基因或位点特异性整合（SSI）靶位点。

2. 发明申请

WO2015026883A1 PLANT GENOME MODIFICATION USING GUIDE RNA/CAS ENDONUCLEASE SYSTEMS AND METHODS OF USE 审中-公开
标题翻译：使用指南RNA / CAS内切酶系统进行植物基因组修饰及其使用方法
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
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
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。

3. 发明申请

WO2013166113A1 COMPOSITIONS AND METHODS COMPRISING SEQUENCES HAVING MEGANUCLEASE ACTIVITY 审中-公开
标题翻译：组合物和包含游离基活性的序列的方法
公开(公告)号：WO2013166113A1
公开(公告)日：2013-11-07
申请号：PCT/US2013/039011
申请日：2013-05-01
申请人： E. I. DU PONT DE NEMOURS AND COMPANY , PIONEER HI-BRED INTERNATIONAL, INC.
发明人： BERMUDEZ, Ericka , CIGAN, Andrew, Mark , ENGLISH, James , FALCO, Saverio, Carl , GAO, Huirong , LIU, Lu , LIU, Zhan-Bin , ONG, Azalea , SVITASHEV, Sergei , YOUNG, Joshua, K.
IPC分类号： C12N9/22 , C12N15/82
CPC分类号： C12N9/22 , C12N9/16 , C12N15/81 , C12N15/8213 , C12Y301/00
摘要： Compositions and methods comprising polynucleotides and polypeptides having meganuclease activity are provided. Further provided are nucleic acid constructs, yeast, plants, plant cells, explants, seeds and grain having the meganuclease sequences. Various methods of employing the meganuclease sequences are provided. Such methods include, for example, methods for producing a meganuclease with increased activity at a wide range of temperatures, methods for producing a yeast, plant, plant cell, explant or seed comprising a meganuclease with increased activity.
摘要翻译：提供了包含具有大范围核酸酶活性的多核苷酸和多肽的组合物和方法。还提供了具有大范围核酸酶序列的核酸构建体，酵母，植物，植物细胞，外植体，种子和颗粒。提供了使用大范围核酸酶序列的各种方法。这样的方法包括例如用于生产在宽温度范围内具有增加的活性的大范围核酸酶的方法，用于生产酵母，植物，植物细胞，外植体或包含具有增加的活性的大范围核酸酶的种子的方法。

4. 发明申请

WO2013019411A1 METHODS AND COMPOSITIONS FOR TARGETED INTEGRATION IN A PLANT 审中-公开
标题翻译：方法和组合在工厂中进行集中整合
公开(公告)号：WO2013019411A1
公开(公告)日：2013-02-07
申请号：PCT/US2012/047202
申请日：2012-07-18
申请人： E. I. DU PONT DE NEMOURS AND COMPANY , PIONEER HI-BRED INTERNATIONAL , 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
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.
摘要翻译：提供组合物和方法用于将目的多核苷酸序列靶向整合到植物或植物细胞的基因组中。方法和组合物使用核酸内切酶和内切核酸酶的识别位点与位点特异性重组位点/重组酶组合，以提供用于在植物，植物细胞或种子的基因组内建立靶位点的有效系统。一旦建立了这样的靶位点，就可以采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。

5. 发明申请

WO2009006297A2 METHODS FOR ALTERING THE GENOME OF A MONOCOT PLANT CELL 审中-公开
标题翻译：改变单核细胞植物细胞基因组的方法
公开(公告)号：WO2009006297A2
公开(公告)日：2009-01-08
申请号：PCT/US2008/068570
申请日：2008-06-27
申请人： PIONEER HI-BRED INTERNATIONAL, INC. , LYZNIK, L., Aleksander , TAO, Yumin , GAO, Huirong
发明人： LYZNIK, L., Aleksander , TAO, Yumin , GAO, Huirong
IPC分类号： C12N15/82 , C12N9/22 , A01H5/00
CPC分类号： C12N15/8201 , A01H1/02 , C12N9/22 , C12N15/8213 , C12N15/8241 , C12Q1/6895 , C12Q2600/13 , C12Q2600/156
摘要： Methods and compositions for altering the genome of a monocot plant cell, and a monocot plant are disclosed. The methods and compositions use a double-strand break inducing agent to alter a monocot plant or plant cell genomic sequence comprising a recognition sequence for the double-strand break inducing agent.
摘要翻译：公开了用于改变单子叶植物细胞和单子叶植物基因组的基因组的方法和组合物。该方法和组合物使用双链断裂诱导剂来改变包含双链断裂诱导剂的识别序列的单子叶植物或植物细胞基因组序列。

6. 发明申请

WO2017132239A1 WAXY CORN 审中-公开
标题翻译：蜡质玉米
公开(公告)号：WO2017132239A1
公开(公告)日：2017-08-03
申请号：PCT/US2017/014903
申请日：2017-01-25
申请人： PIONEER HI-BRED INTERNATIONAL, INC.
发明人： CIGAN, Andrew Mark , GADLAGE, Mark Jacob , GAO, Huirong , MEELEY, Robert B. , YOUNG, Joshua K.
IPC分类号： C12N15/82 , C12N9/10 , A01H5/10
CPC分类号： C12N15/8245 , A01H5/10 , C12N9/1051
摘要： The present disclosure involves the production of Waxy maize. Compositions and methods are provided for knocking out expression of the Waxy (Wx1) gene in maize by making double strand breaks at one or more target sites in an endogenous WX1 encoding sequence. Some methods employ one or more guide polynucleotides and a Cas endonuclease, wherein Cas endonuclease is guided by the one or more guide polynucleotides to recognize and introduce double strand breaks at specific target sites in and around the Wx1 gene. Also provided are compositions and methods for the production of Waxy maize plant cells, plant explants, seeds and grain.
摘要翻译：本公开涉及糯玉米的生产。提供了组合物和方法用于通过在内源性WX1编码序列中的一个或多个靶位点处产生双链断裂来敲除玉米中Waxy（Wx1）基因的表达。一些方法使用一种或多种引导性多核苷酸和Cas内切核酸酶，其中Cas内切核酸酶由一个或多个引导多核苷酸引导以识别并在Wx1基因内和Wx1基因周围的特定靶位点处引入双链断裂。还提供了用于生产糯玉米植物细胞，植物外植体，种子和谷粒的组合物和方法。

7. 发明申请

WO2013066423A2 METHODS AND COMPOSITIONS FOR PRODUCING MALE STERILE PLANTS 审中-公开
标题翻译：用于生产雄性不育植物的方法和组合物
公开(公告)号：WO2013066423A2
公开(公告)日：2013-05-10
申请号：PCT/US2012/043082
申请日：2012-06-19
申请人： PIONEER HI-BRED INTERNATIONAL, INC. , E.I. DU PONT DE NEMOURS AND COMPANY , BIDNEY, Dennis, L. , CIGAN, Mark , FALCO, Carl , GAO, Huirong , JANTZ, Derek , LASSNER, Mike , LOWE, Keith , LYZNIK, Leszek, Aleksander , SMITH, James, Jefferson
发明人： BIDNEY, Dennis, L. , CIGAN, Mark , FALCO, Carl , GAO, Huirong , JANTZ, Derek , LASSNER, Mike , LOWE, Keith , LYZNIK, Leszek, Aleksander , SMITH, James, Jefferson
IPC分类号： C12N15/82
CPC分类号： C12N15/8289 , C12N5/14 , C12N9/22 , C12N15/8213 , C12Q1/6895 , C12Q2600/13 , C12Q2600/156 , C12Q2600/158 , C12Y301/00
摘要： Methods of making a targeted modification in a male fertility gene in the genome of a plant are disclosed. The methods involve contacting a plant cell with an engineered double-strand-break-inducing agent capable of inducing a double-strand break in a target sequence in the male fertility gene and identifying a eel! comprising an alteration in the target sequence. Also disclosed are plants, plant cells, plant parts, and seeds comprising a male fertility gene with an alteration in a male fertility gene. Nucleic acid molecules comprising male fertility genes with at least one targeted modification therein, optimized nucleic acid molecules encoding endonucleases that are engineered double-strand- break-inducing agents and expression cassettes, host cells, and plants comprising one or more of the nucleic acid molecules are further disclosed.
摘要翻译：公开了在植物基因组中对雄性能育基因进行靶向修饰的方法。所述方法包括使植物细胞与能够在雄性能育基因中的靶序列中诱导双链断裂的工程化双链断裂诱导剂接触，并鉴定鳗鱼！包括目标序列的改变。还公开了包含雄性育性基因改变的雄性育性基因的植物，植物细胞，植物部分和种子。包含其中具有至少一种靶向修饰的雄性育性基因的核酸分子，编码作为工程化双链断裂诱导剂和表达盒的核酸内切酶的优化核酸分子，宿主细胞和包含一种或多种核酸分子的植物进一步公开。

8. 发明申请

WO2020131986A1 MULTIPLEX GENOME TARGETING 审中-公开
公开(公告)号：WO2020131986A1
公开(公告)日：2020-06-25
申请号：PCT/US2019/067032
申请日：2019-12-18
申请人： PIONEER HI-BRED INTERNATIONAL, INC.
发明人： GAO, Huirong , YOUNG, Joshua K.
IPC分类号： C12N15/113 , C12N9/22 , C12N15/85
摘要： Methods and compositions are provided for the editing of a plurality of target polynucleotides, for example in the genome of a cell, using a Cas endonuclease guided by a plurality of discrete guide RNAs provided as a poly-guide RNA sequence, wherein each of the discrete guide RNAs are flanked by a sequence (or sequences) capable of recognition and cleavage by an endoribonuclease. Also provided compositions of an endoribonuclease, and methods of use thereof. The endoribonuclease may be obtained or derived from Streptococcus thermophilus.

9. 发明申请

WO2018071362A1 GENERATING NORTHERN LEAF BLIGHT RESISTANT MAIZE 审中-公开
标题翻译：产生北叶抗白粉病玉米
公开(公告)号：WO2018071362A1
公开(公告)日：2018-04-19
申请号：PCT/US2017/055835
申请日：2017-10-10
申请人： PIONEER HI-BRED INTERNATIONAL, INC.
发明人： GAO, Huirong , LI, Bailin , MEELEY, Robert B. , PERUGINI, Leandro Daniel , TABOR, Girma M.
IPC分类号： A01H1/04 , C12Q1/68 , C12N15/82
摘要： Compositions and methods for obtaining plant cells with modified Ht1 nucleotide sequences, modified NLB18 sequences, or both, are provided herein. The methods involve introducing double-strand breaks into the maize genome in an endogenous Ht1 encoding sequence, an endogenous NLB18 encoding sequence, or both, to modify the genomic sequence in order to enhance northern leaf blight resistance of a plant produced from the plant cell. Further provided are methods that introduce resistant alleles of Ht1 and/or NLB18 into specific sites in the genome. Plants produced by the plant cells, and seeds produced from the plants are also included. Guide polynucleotides are also provided for the use of the CRISPR-Cas system in inducing double strand breaks.
摘要翻译：本文提供了用于获得具有修饰的Ht1核苷酸序列，修饰的NLB18序列或两者的植物细胞的组合物和方法。所述方法包括在内源Ht1编码序列，内源NLB18编码序列或两者中将玉米基因组的双链断裂引入玉米基因组中以修饰基因组序列以增强由植物细胞产生的植物的北方叶枯病抗性。进一步提供了将Ht1和/或NLB18的抗性等位基因引入基因组中的特定位点的方法。还包括由植物细胞产生的植物和由植物产生的种子。还提供了使用CRISPR-Cas系统诱导双链断裂的指南多核苷酸。

10. 发明申请

WO2005037863A2 ALTERNATIVE SPLICING FACTORS POLYNUCLEOTIDES, POLYPEPTIDES AND USES THEREOF 审中-公开
标题翻译：替代性分离因子多核苷酸，多肽及其用途
公开(公告)号：WO2005037863A2
公开(公告)日：2005-04-28
申请号：PCT/US2004/031241
申请日：2004-09-23
申请人： PIONEER HI-BRED INTERNATIONAL, INC. , LYZNIK, L. Alexander , GORDON-KAMM, William J. , GAO, Huirong
发明人： LYZNIK, L. Alexander , GORDON-KAMM, William J. , GAO, Huirong
IPC分类号： C07K14/415
CPC分类号： C12N15/827 , C07K14/415 , C12N15/8201 , C12N15/8209 , C12N15/8216 , C12N15/8245 , C12N15/8261 , C12N15/8267 , C12N15/8271 , C12N15/8291 , Y02A40/146
摘要： The invention provides isolated polynucleotides and their encoded proteins that are involved in splicing or modulating splicing activity. The invention further provides recombinant expression cassettes, host cells, transgenic plants, and antibody compositions. The present invention provides methods and compositions relating to altering splicing protein content and/or composition of plants.
摘要翻译：本发明提供分离的多核苷酸及其编码的蛋白质，其涉及剪接或调节剪接活性。本发明还提供了重组表达盒，宿主细胞，转基因植物和抗体组合物。本发明提供了关于改变剪接蛋白质含量和/或植物组成的方法和组合物。

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