E.i. Dupont De Nemours and Company

Pioneer Hi-bred International, Inc.

Corteva Agriscience LLC

Zhan-Bin Liu

Saverio Carl Falco

Andrew Mark Cigan

Huirong Gao

Zhongsen Li

Sergei Svitashev

Michael W Lassner

Anthony John Kinney

Bo Shen

James Jefferson Smith

Derek Jantz

Lu Liu

Joshua K Young

James J English

Ericka Bermudez

Azalea S Ong

Christopher Jay Scelonge

John D Everard

Leszek Aleksander Lyznik

Johan M Stoop

Stephen M Allen

Howard Glenn Damude

William Dean Hitz

Knut Meyer

Edgar Benjamin Cahoon

David J Peterson

Kevin G Ripp

Joan Tellefsen Odell

Jinrui Shi

Thomas G Patterson

Keith Roesler

Kristin Haug Collet

Hyeon-Je Cho

Dean Podlich

Kayla S Flyckt

Laura L Wayne

Jasdeep Mutti

Mark Cigan

Carl Falco

L Aleksander Lyznik

Charles W Kolar, Jr

Qun Zhu

Vesna Djukanovic

Spencer Charles Jones

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.

Methods for producing genetic modifications in a plant genome without incorporating a selectable transgene marker, and compositions thereof

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.

Generation of site-specific-integration sites for complex trait loci in corn and soybean, and methods of use

Provided are compositions comprising polynucleotides encoding modified diacylglycerol acyltransferase-1 (DGAT1) polypeptides having improved properties, such as increased enzymatic activity and/or increased stability. Plants, plant cells, seed, grain and comprising the polynucleotides are provided which have one or more of increased fatty acid or protein content. Methods of generating the polynucleotides in plant cells include transformation and genetic modification. Methods of employing the polynucleotides in plants, methods for increasing DGAT1 activity in a plant, and methods for increasing fatty acid content or protein content in a plant are provided.

Modified seed oil content by gene editing

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.

Methods for producing a complex transgenic trait locus

The disclosure relates to gene expression regulatory sequences, specifically to the promoter of a U6 polymerase III gene and fragments thereof and their use in promoting the expression of one or more heterologous nucleic acid fragments in plants. The invention further discloses compositions, polynucleotide constructs, transformed host cells, mutated plants, transgenic plants and seeds containing the recombinant construct with the promoter, and methods for preparing and using the same.

U6 polymerase III promoter and methods of use

Methods and compositions to modify the fatty acid profiles of seeds and the oils produced therefrom include the modification of four or more alleles of fatty acid desaturases in the genome of a plant. Compositions, polynucleotide constructs, transformed and modified host cells, and plants and seeds exhibiting altered fatty characteristics are provided.

Compositions and methods for modification of fatty acids in soybean

Soybean plants producing soybean seeds comprising leghemoglobin are produced by modifying the genome of the soybean plant. Soybean plants, soybean seeds and soy protein compositions comprising leghemoglobin are provided. Soybean plants, soybean seeds and soy protein compositions comprising leghemoglobin and additionally one or more of high oleic acid, low linolenic acid, high protein, low stachyose, low raffinose and low protease inhibitors are provided. Protein compositions comprising leghemoglobin, such as soy isolates and concentrates can be made from the soybean seeds. Additionally, methods for generating and using plants, seeds and protein compositions comprising leghemoglobin are disclosed.

Leghemoglobin in soybean

Soybean seeds with increased protein, oil or protein plus oil and having a modified expression of a reticulon-like polypeptide, modified expression of a trehalose-6-phosphate synthase polypeptide, or modified expression of both sequences are provided. Methods for modifying expression of reticulon-like polypeptides and polynucleotides and trehalose-6-phosphate synthase polypeptides and polynucleotides include genome editing to modify the transcription regulatory region or sequence encoding the reticulon-like and trehalose-6-phosphate synthase polypeptides and transformation with recombinant DNA constructs to enhance or suppress expression.

Modified genes to increase seed protein content

The disclosure relates to gene expression regulatory sequences from maize, specifically to the promoter of a maize U6 polymerase III gene and fragments thereof and their use in promoting the expression of one or more heterologous nucleic acid fragments in plants. The invention further discloses compositions, polynucleotide constructs, transformed host cells, mutated plants, transgenic plants and seeds containing the recombinant construct with the promoter, and methods for preparing and using the same.

The present disclosure includes the production of a mutant plant resistant to an herbicide of the phosphonomethylglycine family, e.g. glyphosate. Compositions and methods are provided for editing a nucleotide sequence of interest in a cell employing a guide polynucleotide/Cas endonuclease system, wherein the Cas endonuclease is guided by a guide polynucleotide to recognize and optionally introduce a double strand break at a specific target site into the genome of a cell. The nucleotide sequence of interest to be edited can be located within or outside the target site that is recognized by a Cas endonuclease. More specifically, compositions and methods are provided for editing an enolpyruvylshikimate-3-phosphate synthase (EPSPS) nucleotide sequence in a cell. The methods and compositions employ a guide polynucleotide/Cas endonuclease system to provide for an effective system for editing EPSPS nucleotide sequences of within the genome of a cell. Also provided are compositions and methods for the production of glyphosate tolerant plant cells, plants explants, seeds and grain.

Compositions and methods for producing plants resistant to glyphosate herbicide

The disclosure relates to gene expression regulatory sequences from soybean, specifically to the promoter of a soybean U6 polymerase III gene and fragments thereof and their use in promoting the expression of one or more heterologous nucleic acid fragments in plants. The invention further discloses compositions, polynucleotide constructs, transformed host cells, mutated plants, transgenic plants and seeds containing the recombinant construct with the promoter, and methods for preparing and using the same.

Soybean U6 polymerase III promoter and methods of use

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.

Compositions and methods comprising sequences having meganuclease activity

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.

Methods and compositions for targeted integration in a plant

Growing community of inventors

West Chester, PA, United States of America

Zhan-Bin Liu