Agronomic trait modification using guide rna/cas endonuclease systems and methods of use
AssigneePioneer Hi-Bred International, Inc.
InventorsHUIRONG GAO, XIAOMU NIU, JINRUI SHI
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Abstract
Compositions and methods are provided for agronomic trait 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 a genomic region of a plant, plant cell or seed to provide improvement in a desirable agronomic trait such as drought, yield, and stress tolerance. Breeding 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.
Claims51
A method of improving an agronomic trait of a plant, the method comprising providing a guide RNA that targets a polynucleotide involved in improving one or more agronomic characteristics of the plant in association with a Cas endonuclease that creates a double strand break at the polynucleotide and generating the plant, wherein the plant exhibits an improvement in the agronomic trait.
A method of improving grain yield of a maize plant, the method comprising providing a guide RNA that targets a polynucleotide involved in ethylene biosynthesis or ethylene signaling, the guide RNA acts in association with a Cas endonuclease that creates a double strand break at the polynucleotide and generating the plant, wherein the maize plant exhibits improved grain yield.
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A method for editing a regulatory sequence involved in the expression of a gene for abiotic stress tolerance, in the genome of a cell, the method comprising introducing a guide polynucleotide, a polynucleotide modification template and at least one Cas endonuclease into a cell, wherein said guide RNA and Cas endonuclease are capable of forming a complex that enables the Cas endonuclease to introduce a double strand break at a target site in the genome of said cell, wherein said polynucleotide modification template comprises at least one nucleotide modification of said nucleotide sequence.
A method for replacing a first regulatory sequence modulating the expression of a gene involved in agronomic trait in a cell, the method comprising introducing a guide RNA, a polynucleotide modification template, and a Cas endonuclease into said cell, wherein said guide RNA and Cas endonuclease are capable of forming a complex that enables the Cas endonuclease to introduce a double strand break at a target site in the genome of said cell, wherein said polynucleotide modification template comprises a second promoter or second promoter fragment that is different from said first promoter sequence.
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Dependencies are inferred from the claim text.