Methodologies and compositions for creating targeted recombination and breaking linkage between traits
Inventors
Kumar, Sandeep • Lee, Ryan M. • Novak, Stephen • Worden, Andrew F.
Assignees
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Publication Number
US-12590313-B2
Publication Date
2026-03-31
Expiration Date
Abstract
The current disclosure relates to methods and compositions for improving plant varieties through plant breeding and plant genetics. For instance, the disclosure concerns increasing the recombination frequency of a heterozygous trait genetically linked to a second trait within plants. Further, the disclosure concerns breaking the genetic linkage between a first allele and a second allele.
Core Innovation
The disclosure describes a method for increasing the frequency of genetic recombination between a first locus genetically linked to a second locus within a genome of a plant. The method introduces a site specific nuclease into the plant genome, produces a double stranded break at a genomic target site with the nuclease in one of two homologous chromosomes, and does not produce the double stranded break in the second homologous chromosome.
The method then undergoes recombination within the plant genome and modifies the plant genome to comprise an increased frequency of genetic recombination between the first locus and the second locus. The disclosed approach is directed to reducing genetic linkage between the first locus and the second locus by increasing recombination frequency while breaking linkage drag between genetically linked loci.
The recombination is described in contexts that include meiotic recombination and mitotic recombination, and the loci are described as first and second loci that are genetically linked. The disclosure further specifies implementation aspects for the site specific nuclease and its delivery, including zinc finger nucleases, TALEN nucleases, CRISPR nucleases, meganucleases, leucine zipper nucleases, intragenomic delivery, and direct delivery.
Claims Coverage
The independent claim establishes a method to increase genetic recombination between genetically linked loci by introducing a site-specific nuclease that creates a double stranded break in only one homolog, followed by recombination and genome modification to reduce linkage. The dependent claims add quantified recombination increase, locus-distance ranges, nuclease type selection, nuclease delivery modes, and enumerated plant species.
Nuclease-mediated single-homolog double-strand break for recombination between linked loci
A method for increasing the frequency of genetic recombination between a first locus genetically linked to a second locus within a genome of a plant by introducing a site specific nuclease into the genome of the plant; producing a double stranded break at a genomic target site with the site specific nuclease in one of two homologous chromosomes where a double stranded break is not produced in the second homologous chromosome; undergoing recombination within the plant genome; and modifying the plant genome to comprise increased frequency of genetic recombination between the first locus and the second locus to reduce genetic linkage between the first locus and the second locus.
Quantified recombination increase
The method results in an increased frequency of genetic recombination ranging from 1.25 to 17.8 fold.
Locus distance constrained in physical and genetic distance
The distance from the first locus to the second locus ranges from about 10 bp to about 10 Mbp.
Site-specific nuclease selected from specified nuclease classes
The site specific nuclease is selected from the group consisting of a zinc finger nuclease, a TALEN nuclease, a CRISPR nuclease, a meganuclease, and a leucine zipper nuclease.
Nuclease delivery by intragenomic recombination or direct delivery
The site specific nuclease is delivered to a cell by intragenomic recombination or via direct delivery.
Plant selected from an enumerated species set
The plant is selected from the group consisting of a tobacco plant, a soybean plant, a cotton plant, a Brassica plant, a corn plant, a sorghum plant, a wheat plant, and a rice plant.
Across the independent and dependent claims, the core claim covers increasing recombination between genetically linked loci by creating a double stranded break in only one homolog using a site specific nuclease, followed by recombination and genome modification to reduce linkage. The dependent claims further constrain the approach with quantified recombination increase, locus-distance ranges, selectable nuclease classes, specified delivery modes, and enumerated plant species.
Stated Advantages
Increase the frequency of genetic recombination between the first locus and the second locus to reduce genetic linkage between the first locus and the second locus.
Produce a modified plant genome with reduced genetic linkage between the first locus and the second locus.
Documented Applications
No documented applications found
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