# US 8,541,655 B2 · Transgenic plants expressing modified Cry3A

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- Number: US 8,541,655 B2
- Document ID: US8541655B2
- Status: Granted
- Kind: B2
- Application number: 13/525,413
- Filed: 2012-06-18
- Published: 2012-10-11
- Granted: 2013-09-24
- Earliest claimed: 2001-08-31
- Assignees: Syngenta Participations AG
- Applicants: Eric Chen; Cheryl Stacy
- Inventors: Eric Chen; Cheryl Stacy
- Web page: https://patentgrep.com/patent/US8541655B2

## CPC

- C12N 15/8286: {for insect resistance}
- C07K 14/325: Bacillus thuringiensis crystal peptides, i.e. delta-endotoxins
- Y02A 40/146: Genetically Modified [GMO] plants, e.g. transgenic plants
- C12N: MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA (microbiological testing media C12Q1/00)
- 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)
- Y02A: TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE

## Abstract

Compositions and methods for controlling plant pests are disclosed. In particular, transgenic maize plants comprising nucleic acid molecules encoding modified Cry3A toxins having increased toxicity to western corn rootworm are provided. Such modified Cry3A toxins, into which a non-naturally occurring protease recognition site is inserted, are expressed at levels in transgenic plants to inhibit insect feeding damage and cause insect mortality.

## Claims

### Claim 1

A transgenic plant comprising an isolated nucleic acid molecule that encodes a modified Cry3A toxin comprising a non-naturally occurring protease recognition site, wherein said protease recognition site modifies a Cry3A toxin and is located at a position between amino acids corresponding to amino acid numbers 107 and 113 of SEQ ID NO:4, wherein said protease recognition site is recognizable by a gut protease of western corn rootworm, and wherein said modified Cry3A toxin causes higher mortality to western corn rootworm than the mortality caused by said Cry3A toxin to western corn rootworm in an artificial diet bioassay.

### Claim 2 (depends on 1)

The transgenic plant according to claim 1, wherein said gut protease is a serine protease or a cysteine protease.

### Claim 3 (depends on 1)

The transgenic plant according to claim 1, wherein the protease recognition site is located at a position corresponding to amino acid numbers 107 and 111 of SEQ ID NO:4.

### Claim 4 (depends on 1)

The transgenic plant according to claim 1, wherein said protease recognition site is located between amino acid numbers 107 and 113 of SEQ ID NO:4.

### Claim 5 (depends on 4)

The transgenic plant according to claim 4, wherein said protease recognition site is located between amino acid numbers 107 and 111 of SEQ ID NO:4.

### Claim 6 (depends on 1)

The transgenic plant according to claim 1, wherein the nucleic acid molecule is operably linked to a heterologous promoter sequence.

### Claim 7 (depends on 1, 2, 3, 4, 5, 6)

The transgenic plant according to any of claims 1 to 6, wherein said plant is a maize plant.

### Claim 8 (depends on 1)

Transgenic seed from the transgenic plant of claim 1, wherein said seed comprises the nucleic acid molecule.

### Claim 9 (depends on 7)

Transgenic seed from the maize plant of claim 7, wherein said seed comprises the nucleic acid molecule.

### Claim 10 (depends on 7)

The transgenic maize plant according to claim 7, wherein said maize plant expresses said modified Cry3A toxin in root tissue at a level that causes mortality to at least western corn rootworm.

### Claim 11 (depends on 10)

The transgenic maize plant according to claim 10, which is an inbred maize plant.

### Claim 12 (depends on 10)

The transgenic maize plant according to claim 10, which is a hybrid maize plan.

### Claim 13 (depends on 11)

Transgenic seed of the transgenic inbred maize plant according to claim 11, wherein said seed comprises the nucleic acid molecule.

### Claim 14 (depends on 12)

Transgenic seed of the transgenic hybrid maize plant according to claim 12, wherein said seed comprises the nucleic acid molecule.