# US 2023/0057587 A1 · Shatterproof genes and mutations

> Agents: this data is available as JSON from the patentgrep API and MCP server — see https://patentgrep.com/llms.txt

- Number: US 2023/0057587 A1
- Document ID: US20230057587A1
- Status: Published application
- Kind: A1
- Application number: 17/744,422
- Filed: 2022-05-13
- Published: 2023-02-23
- Earliest claimed: 2018-01-09
- Assignees: Cibus US LLC; Cibus Europe B.V.
- Applicants: Cibus US LLC; Cibus Europe, B.V.
- Inventors: Gregory F.W. GOCAL
- Web page: https://patentgrep.com/patent/US20230057587A1

## CPC

- C07K 14/415: from plants
- C12N 15/8266: {Abscission; Dehiscence; Senescence}
- C12N 15/8213: {Targeted insertion of genes into the plant genome by homologous recombination}
- C07K: PEPTIDES (peptides in foodstuffs A23; obtaining protein compositions for foodstuffs, working-up proteins for foodstuffs A23J; preparations for medicinal purposes A61K; peptides containing beta-lactam rings C07D; cyclic dipeptides not having in their molecule any other peptide link than those which form their ring, e.g. piperazine-2,5-diones, C07D; ergot alkaloids of the cyclic peptide type C07D519/02; macromolecular compounds having statistically distributed amino acid units in their molecules, i.e. when the preparation does not provide for a specific; but for a random sequence of the amino acid units, homopolyamides and block copolyamides derived from amino acids C08G69/00; macromolecular products derived from proteins C08H1/00; preparation of glue or gelatine C09H; single cell proteins, enzymes C12N; genetic engineering processes for obtaining peptides C12N15/00; compositions for measuring or testing processes involving enzymes C12Q; investigation or analysis of biological material G01N33/00)
- C12N: MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA (microbiological testing media C12Q1/00)

## Abstract

The present disclosure provides shatterproof (SHP) genes and plants and/or plant cells bearing one or more mutations in a shatterproof gene; as well as methods of making and using such plants. In some embodiments the plant or plant cell is resistant to preharvest dehiscence.

## Claims

### Claim 1

(canceled)

### Claim 2

(canceled)

### Claim 3

(canceled)

### Claim 4

(canceled)

### Claim 5

(canceled)

### Claim 6

(canceled)

### Claim 7

(canceled)

### Claim 8

(canceled)

### Claim 9

(canceled)

### Claim 10

(canceled)

### Claim 11

(canceled)

### Claim 12

(canceled)

### Claim 13

(canceled)

### Claim 14

(canceled)

### Claim 15

(canceled)

### Claim 16

(canceled)

### Claim 17

(canceled)

### Claim 18

(canceled)

### Claim 19

(canceled)

### Claim 20

(canceled)

### Claim 21

(canceled)

### Claim 22

(canceled)

### Claim 23

(canceled)

### Claim 24

(canceled)

### Claim 25

(canceled)

### Claim 26

(canceled)

### Claim 27

(canceled)

### Claim 28

(canceled)

### Claim 29

(canceled)

### Claim 30

(canceled)

### Claim 31

(canceled)

### Claim 32

(canceled)

### Claim 33

(canceled)

### Claim 34

(canceled)

### Claim 35

(canceled)

### Claim 36

(canceled)

### Claim 37

(canceled)

### Claim 38

(canceled)

### Claim 39

(canceled)

### Claim 40

(canceled)

### Claim 41

(canceled)

### Claim 42

(canceled)

### Claim 43

(canceled)

### Claim 44

(canceled)

### Claim 45

(canceled)

### Claim 46

(canceled)

### Claim 47

A plant or part thereof comprising at least one mutation in at least three, at least four, at least five, at least six, at least seven, or eight nucleic acid sequences encoding SHATTERPROOF (SHP) genes.

### Claim 48 (depends on 47)

The plant or part thereof of claim 47, wherein the nucleic acid sequences have at least 90% sequence identity, at least 95% sequence identity, at least 98% sequence identity, or at least 99% sequence identity to nucleic acid sequences selected from the group consisting of SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3, SEQ ID NO: 4, SEQ ID NO: 5, SEQ ID NO: 6, SEQ ID NO: 7, and SEQ ID NO: 8.

### Claim 49 (depends on 47)

The plant or part thereof of claim 47, wherein the nucleic acid sequences are selected from the group consisting of SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3, SEQ ID NO: 4, SEQ ID NO: 5, SEQ ID NO: 6, SEQ ID NO: 7, and SEQ ID NO: 8.

### Claim 50 (depends on 47)

The plant or part thereof of claim 47, wherein the mutation is a frameshift mutation.

### Claim 51 (depends on 50)

The plant or part thereof of claim 50, wherein the frameshift mutation results in one or more nucleotide insertions or deletions as compared to the corresponding endogenous gene without the frameshift mutation.

### Claim 52 (depends on 50)

The plant or part thereof of claim 50, wherein the frameshift mutation results in a premature stop codon.

### Claim 53 (depends on 50)

The plant or part thereof of claim 50, wherein the mutation reduces or eliminates the expression or activity of the SHP gene and/or SHP polypeptide.

### Claim 54 (depends on 47)

The plant or part thereof of claim 47, wherein the plant exhibits reduced susceptibility to preharvest dehiscence.

### Claim 55 (depends on 47)

The plant of claim 47, wherein the plant is selected from the group consisting of Brassica napus, Brassica rapa, Brassica oleracea, Brassica juncea, Brassica species, Raphanus sativus, Pisum sativum, Phaseolus vulgaris, Lens culinaris, Glycine max, and Fabaceae species.

### Claim 56 (depends on 47)

A method of producing the plant of claim 47, comprising the steps of:
a) introducing mutations into plant cells, wherein the mutations are at least one mutation in at least three, at least four, at least five, at least six, at least seven, or eight nucleic acid sequences encoding SHP genes;
b) selecting plant cells containing the mutations; and
c) regenerating a plant having the mutations;
wherein the plant exhibits reduced susceptibility to preharvest dehiscence.

### Claim 57 (depends on 56)

The method of claim 56, wherein the mutations are introduced using one or more vectors, wherein the vectors comprise gene editing components selected from the group consisting of a nuclease, an RNA-guided DNA endonuclease, a CRISPR/Cas9 system, a TALEN, a zinc finger, and a meganuclease designed to target a nucleic acid sequence encoding a SHP gene.

### Claim 58 (depends on 56)

The method of claim 56, wherein the mutations are introduced using a GRON system designed to target a nucleic acid sequence encoding a SHP gene.

### Claim 59 (depends on 58)

The method of claim 58, wherein the GRON system comprises one or more modifications selected from the group consisting of a Cy3 group, 3PS group, and a 2′O-methyl group.

### Claim 60 (depends on 56)

The method of claim 56, wherein the nucleic acid sequences have at least 90% sequence identity, at least 95% sequence identity, at least 98% sequence identity, or at least 99% sequence identity to nucleic acid sequences selected from the group consisting of SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3, SEQ ID NO: 4, SEQ ID NO: 5, SEQ ID NO: 6, SEQ ID NO: 7, and SEQ ID NO: 8.

### Claim 61 (depends on 56)

The method of claim 56, wherein the nucleic acid sequences are selected from the group consisting of SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3, SEQ ID NO: 4, SEQ ID NO: 5, SEQ ID NO: 6, SEQ ID NO: 7, and SEQ ID NO: 8.

### Claim 62 (depends on 56)

The method of claim 56, wherein the mutation is selected from the group consisting of a frameshift mutation, a frameshift mutation resulting in one or more nucleotide insertions or deletions as compared to the corresponding endogenous gene without the frameshift mutation, and a frameshift mutation resulting in a premature stop codon, and wherein the mutation reduces or eliminates expression of the SHP gene and/or SHP polypeptide.

### Claim 63 (depends on 56)

The method of claim 56, wherein the plant is selected from the group consisting of Brassica napus, Brassica rapa, Brassica oleracea, Brassica juncea, Brassica species, Raphanus sativus, Pisum sativum, Phaseolus vulgaris, Lens culinaris, Glycine max, and Fabaceae species.

### Claim 64 (depends on 47)

An F1 plant, wherein the F1 plant has the plant of claim 47 as a parent.

### Claim 65 (depends on 47)

A method of making plant seeds, the method comprising crossing the plant of claim 47 with another plant and harvesting seed therefrom.

### Claim 66

(canceled)

### Claim 67 (depends on 65)

A plant produced by growing the seed of claim 65, wherein the plant has all the physiological and morphological characteristics of a plant comprising at least one mutation in at least three, at least four, at least five, at least six, at least seven, or eight nucleic acid sequences encoding SHATTERPROOF (SHP) genes.