US 2026/0265768 A1 · COMPOSITIONS AND METHODS FOR TARGETABLE GENETIC RECOMBINATION IN PLANTS

Abstract

Predictable and reliable gene stacking will simplify the breeding process, saving time and resources. The Cre-Lox recombination system offers the ability to easily add, remove, or alter traits for more precise control over plant characteristics. The system uses specific bespoke loxP sites to ensure unidirectional gene insertion, creating a “Loxbox” to prevent further insertions at the same site. This method allows for iterative, targeted gene stacking approach to user-defined genetic engineering by recombination in a plant cell, plant tissue, plant material, plant part, and/or plant with predictable directionality. By creating a toolkit for targeted sequential gene insertion, the trait development pipeline can be streamlined, making it easier and faster to develop new plant varieties with desirable traits.

Claims20

Dependent claims

A modified plant genome comprising a mutant loxP landing pad nucleotide sequence, wherein the mutant loxP landing pad comprises

(i) a wild-type loxP nucleotide sequence,
optionally wherein the wild-type loxP comprises SEQ ID NO:1 or SEQ ID NO:2;
and
(ii) a first mutant loxP nucleotide sequence;
optionally wherein the first mutant loxP nucleotide sequence comprises a nucleic acid sequence set forth in any one of SEQ ID NOs:
3-14 or 43-46.

3 more elements

A method for iterative stacking of donor genes within a target plant genome comprising

(a) contacting a target plant genome comprising a first mutant loxP landing pad with a first recombination mediated exchange cassette (RMEC), comprising
(i) a wild-type loxP nucleotide sequence,
optionally wherein the wild-type loxP comprises SEQ ID NO:1 or SEQ ID NO:2;
(ii) a second mutant loxP nucleotide sequence;
(iii) a donor nucleotide sequence;
and
(iv) a third mutant loxP nucleotide sequence;
wherein the second and third mutant loxP nucleotide sequence(s) are orientated either side of the donor nucleotide sequence within the RMEC;
and
wherein the wild-type loxP nucleotide sequence and the second mutant loxP nucleotide sequence are orientated upstream of the donor nucleotide sequence within the RMEC;
and
wherein the third mutant loxP sequence is configured to direct Cre recombinase-mediated recombination with the first mutant loxP sequence within the first landing pad,
wherein the contacting occurs under conditions that enable Cre-recombinase mediated exchange of the RMEC into the first landing pad to provide a second mutant loxP landing pad comprising the second mutant loxP sequence;
and
(b) optionally confirming recombination of the donor nucleotide sequence into the target genome.

12 more elements

Dependencies are inferred from the claim text.

Facts

Application

19/557,016Filed 4 Mar 2026

Priority

4 Mar 2025

Published

10 Sep 2026

Term ends

Not estimatedThe source records don’t support an estimate.

Classification8 codes

C12N 15/8216
For plant cells , e.g. plant artificial chromosomes: methods for controlling, regulating or enhancing expression of transgenes in plant cells
C12N 5/04
Undifferentiated human, animal or plant cells, e.g. cell lines; Tissues; Cultivation or maintenance thereof; Culture media therefor: plant cells…
C12N 9/1241
Transferring phosphorus containing groups, e.g. kinases: nucleotidyltransferases
C12N 9/22
Acting on ester bonds: ribonucleases [RNase]; Deoxyribonucleases [DNase]

Compositions and methods for targetable genetic recombination in plants

Life of this patent

Compositions and methods for targetable genetic recombination in plants

Life of this patent

Abstract

Claims20