# US 2021/0274783 A1 · Engineered pesticidal proteins and methods of controlling plant pests > Agents: this data is available as JSON from the patentgrep API and MCP server — see https://patentgrep.com/llms.txt - Number: US 2021/0274783 A1 - Document ID: US20210274783A1 - Status: Published application - Kind: A1 - Application number: 16/468,528 - Filed: 2017-11-29 - Published: 2021-09-09 - Earliest claimed: 2016-12-12 - Assignees: Syngenta Participations AG - Applicants: Syngenta Participations AG - Inventors: Hyunsook Sarah Chae - Web page: https://patentgrep.com/patent/US20210274783A1 ## CPC - C12N 15/8286: {for insect resistance} - A01N 37/46: N-acyl derivatives - A01N 63/50: Isolated enzymes; Isolated proteins (peptides A01N37/46) - 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) - A01N: PRESERVATION OF BODIES OF HUMANS OR ANIMALS OR PLANTS OR PARTS THEREOF (preservation of food or foodstuff A23); BIOCIDES, e.g. AS DISINFECTANTS, AS PESTICIDES OR AS HERBICIDES (preparations for medical, dental or toiletry purposes which kill or prevent the growth or proliferation of unwanted organisms A61K); PEST REPELLANTS OR ATTRACTANTS; PLANT GROWTH REGULATORS - 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 The invention provides nucleic acids, polypeptides, transgenic plants, compositions and methods for conferring pesticidal activity (e.g., insecticidal activity) to bacteria, plants, plant cells, tissues and seeds. Nucleic acids encoding the insecticidal proteins can be used to transform prokaryotic and eukaryotic organisms to express the insecticidal proteins. The recombinant organisms or compositions containing the recombinant organisms or insecticidal proteins or in combination with an appropriate agricultural carrier can be used to control an insect pest in various environments. ## Claims ### Claim 1 A chimeric insecticidal protein that is toxic to a lepidopteran insect pest comprising in an N-terminal to C-terminal direction: (a) an N-terminal region of a first Cry1 protein, which is an N-terminal region of a BT-0029 protein of SEQ ID NO:2 or an amino acid sequence that is at least 90% identical thereto, fused to (b) a C-terminal region of a different Cry1 protein; wherein a crossover position between the first Cry1 protein and the different Cry1 protein is located in conserved block 3. ### Claim 2 (depends on 1) The chimeric insecticidal protein according to claim 1, wherein the different Cry1 protein is a) a Cry1F, a Cry1G, a Cry1I, a Cry1K or a Cry1C protein; or b) is a Cry1Fa, a Cry1Ia, a Cry1If, a Cry1Ka or a Cry1Ca; or c) is a BT-0022 protein of SEQ ID NO:1. ### Claim 3 (depends on 1) The chimeric insecticidal protein according to claim 1, wherein the C-terminal region of the different Cry1 protein comprises an amino acid sequence corresponding to an amino acid sequence from a) position 464 to 602 of SEQ ID NO: 8 or an amino acid sequence that is at least 80% identical thereto; or b) position 498 to 636 of SEQ ID NO: 1 or an amino acid sequence that is at least 80% identical thereto. ### Claim 4 (depends on 1) The chimeric insecticidal protein according to claim 1, wherein the N-terminal region of the first Cry1 protein comprises an amino acid sequence corresponding to an amino acid sequence from position 1 to 458 of SEQ ID NO: 2 or an amino acid sequence that is at least 90% identical thereto. ### Claim 5 (depends on 1) The chimeric insecticidal protein according to claim 1, wherein the chimeric insecticidal protein further comprises at the C-terminus a pro-toxin tail from a Cry protein that is cleaved from the protoxin after ingestion by the lepidopteran insect pest. ### Claim 6 (depends on 1) The chimeric insecticidal protein according to claim 1, wherein the chimeric insecticidal protein comprises: (a) the amino acid sequence from position 1 to 597 of SEQ ID NO: 3 or a toxin fragment thereof, or (b) the amino acid sequence of SEQ ID NO: 3 or a toxin fragment thereof; or (c) the amino acid sequence from positions 1 to 597 of SEQ ID NO: 9 or a toxin fragment thereof; or (d) the amino acid sequence of SEQ ID NO: 9 or a toxin fragment thereof; or (e) an amino acid sequence that is at least 80% identical to the amino acid sequence of (a), (b), (c) or (d). ### Claim 7 (depends on 1) The chimeric insecticidal protein according to claim 1, wherein the chimeric insecticidal protein has insecticidal activity against a Spodoptera frugiperda insect pest with resistance to a Vip3A protein or a Cry1F protein. ### Claim 8 (depends on 1) A polynucleotide comprising (a) a nucleotide sequence encoding the chimeric insecticidal protein according to claim 1; or (b) the nucleotide sequence of (a) that is codon optimized for expression in a plant. ### Claim 9 (depends on 8) The polynucleotide according to claim 8, wherein the polynucleotide comprises a nucleotide sequence that comprises: (a) a nucleotide sequence of SEQ ID NO: 4, SEQ ID NO: 5, SEQ ID NO: 6, SEQ ID NO: 7, SEQ ID NO: 10, SEQ ID NO: 11 or SEQ ID NO: 14 or a toxin-encoding fragment thereof; (b) a nucleotide sequence that is substantially identical to the nucleotide sequence of (a); (c) a nucleotide sequence that anneals under stringent hybridization conditions to the nucleotide sequence of (a) or (b); or (d) a nucleotide sequence that differs from the nucleotide sequence of (a), (b) or (c) due to the degeneracy of the genetic code. ### Claim 10 (depends on 8) A nucleic acid molecule comprising the polynucleotide according to claim 8 operably associated with a heterologous plant expressible promoter. ### Claim 11 (depends on 10) A vector comprising the nucleic acid molecule according to claim 10. ### Claim 12 (depends on 8) A transgenic cell comprising the polynucleotide according to claim 8, wherein the transgenic cell is a bacterial cell or a plant cell. ### Claim 13 (canceled) ### Claim 14 (depends on 12) A transgenic plant or part thereof comprising the transgenic plant cell according to claim 12, wherein the plant is: (a) a monocot plant, optionally a barley plant, a maize plant, an oat plant, a rice plant, a sorghum plant, a sugarcane plant or a wheat plant; or (b) a dicot plant, optionally a soybean plant, a sunflower plant, a tomato plant, a cole crop plant, a cotton plant, a sugar beet plant or a tobacco plant. ### Claim 15 (depends on 14) The transgenic plant or part thereof according to claim 14, wherein the transgenic plant further comprises a nucleotide sequence encoding a second insect control agent. ### Claim 16 (canceled) ### Claim 17 (depends on 14) A transgenic seed of the transgenic plant according to claim 14, wherein the seed comprises the polynucleotide. ### Claim 18 (depends on 14) A harvested product derived from the transgenic plant according to claim 14, wherein the harvested product comprises the chimeric insecticidal protein. ### Claim 19 (depends on 18) A processed product derived from the harvested product according to claim 18, wherein the processed product is a flour, a meal, an oil, a starch, or a product derived from any of the foregoing. ### Claim 20 (canceled) ### Claim 21 (depends on 8) A method of producing a transgenic plant with increased resistance to a lepidopteran insect pest, the method comprising: (a) introducing into a plant by transforming a plant cell with the polynucleotide of claim 8; or crossing a first plant comprising the polynucleotide, with a second plant, wherein the chimeric insecticidal protein is expressed in the plant, thereby producing a transgenic plant with increased resistance to an insect pest; and optionally obtaining a progeny plant from the transgenic plant, wherein the progeny plant comprises the polynucleotide and has increased resistance to an insect pest; or (b) planting a seed comprising the polynucleotide and growing a transgenic plant from the seed, wherein the transgenic plant comprises the polynucleotide and produces the chimeric insecticidal protein. ### Claim at position 22 -24. (canceled) ### Claim 25 (depends on 1) A method of controlling a lepidopteran insect pest, the method comprising delivering to the insect pest or an environment thereof a composition comprising an effective amount of the chimeric insecticidal protein of claim 1, wherein the lepidopteran insect pest is susceptible or resistant to a Vip3 protein or a Cry1F protein. ### Claim 26 (depends on 8) A method of reducing the development of resistance to a Vip3A protein or a Cry1F protein in a population of a target lepidopteran insect pest, the method comprising delivering to the target population or an environment thereof a transgenic plant comprising: (i) the polynucleotide according to claim 8; and (ii) a polynucleotide comprising a nucleotide sequence encoding a Vip3A protein or a nucleotide sequence encoding a Cry1F protein; wherein the chimeric insecticidal protein and the Vip3A protein or the Cry1F protein are produced in the transgenic plant.