Granted Patent B2

US 12,593,765 · App. 16/465,644 · Granted Apr 7, 2026

Inventors: Qiudeng Que (Research Triangle Park, NC); Timothy Kelliher (Research Triangle Park, NC)

Assignee: SYNGENTA CROP PROTECTION AG

A01H1/08C07K14/415C12N9/22C12N15/11C12N15/8213C12N15/8216C12N15/8238C12N15/8241C12N15/8287C12Q1/6895C12N2310/20C12N2800/80C12Y301/01032

Abstract

Claims (13)

1 . A method of editing rice plant genomic DNA, comprising:

a) obtaining a first rice plant, wherein the first rice plant comprises a loss-of function mutation in a wildtype patatin-like phospholipase A2α gene encoding a polypeptide having at least 95% identity to SEQ ID NO:118; and the loss-of function mutation is a frameshift mutation or insertion or deletion mutation that prevents expression of an active protein product from the wildtype patatin-like phospholipase A2α gene; and wherein said first rice plant expresses a DNA modification enzyme and optionally at least one guide nucleic acid that targets the genome of a second rice plant at a site to be edited;

b) pollinating the second rice plant with pollen from the first rice plant; and

c) selecting at least one haploid progeny produced by the pollination of step (b) wherein the haploid progeny comprises the genome of the second rice plant but not the first rice plant, and the genome of the haploid progeny has been modified by the DNA modification enzyme and optional at least one guide nucleic acid delivered by the first rice plant.

2 . The method of claim 1 , wherein the DNA modification enzyme is a site-directed nuclease selected from the group consisting of meganucleases (MNs), zinc-finger nucleases (ZFNs), transcription-activator like effector nucleases (TALENs), Cas9 nuclease, Cpf1 nuclease, dCas9-FokI, dCpf1-FokI, chimeric Cas9-cytidine deaminase, chimeric Cas9-adenine deaminase, chimeric FEN1-FokI, Mega-TALs, nickase Cas9 (nCas9), chimeric dCas9 non-FokI nuclease, and dCpf1 non-FokI nuclease.

3 . The method of claim 1 , wherein the at least one guide nucleic acid is a guide RNA.

4 . The method of claim 1 , wherein the edited haploid progeny is treated with a chromosome doubling agent, thereby creating an edited doubled haploid progeny.

5 . The method of claim 4 , wherein the chromosome doubling agent is colchicine, pronamide, dithipyr, or trifluralin.

6 . The method of claim 1 , wherein the first rice plant expresses a marker gene.

7 . The method of claim 6 , wherein the marker gene is selected from the group consisting of GUS, PMI, PAT, GFP, RFP, CFP, B1, C1, and R-nj; or is an anthocyanin pigment.

8 . The method of claim 1 , wherein the wildtype patatin-like phospholipase A2α gene comprises SEQ ID NO:33.

9 . The method of claim 1 , wherein the wildtype patatin-like phospholipase A2α gene encodes SEQ ID NO:118.

10 . The method of claim 1 , wherein the wildtype patatin-like phospholipase A2α gene encodes SEQ ID NO:119.

Assignments (2)

Continuity (2)

References Cited (89)