# US 11,439,152 B2 · Control of hemipteran pests using RNA molecules

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- Number: US 11,439,152 B2
- Document ID: US11439152B2
- Status: Granted
- Kind: B2
- Application number: 16/757,106
- Filed: 2018-10-16
- Published: 2020-07-30
- Granted: 2022-09-13
- Earliest claimed: 2017-10-18
- Assignees: Syngenta Participations AG
- Applicants: Syngenta Participations AG
- Inventors: Lies Degrave; Myriam Beghyn; Yann Naudet; Lien De Schrijver; Kevin V Donohue
- Web page: https://patentgrep.com/patent/US11439152B2

## CPC

- A01N 63/60: Isolated nucleic acids
- A01N 57/16: containing heterocyclic radicals
- C07K 14/195: from bacteria
- C07K 14/24: from Enterobacteriaceae (F), e.g. Citrobacter, Serratia, Proteus, Providencia, Morganella, Yersinia
- C07K 14/325: Bacillus thuringiensis crystal peptides, i.e. delta-endotoxins
- C07K 14/33: from Clostridium (G)
- C07K 14/43563: {from insects}
- C12N 15/113: Non-coding nucleic acids modulating the expression of genes, e.g. antisense oligonucleotides; {Antisense DNA or RNA; Triplex- forming oligonucleotides; Catalytic nucleic acids, e.g. ribozymes; Nucleic acids used in co-suppression or gene silencing (when used in plants C12N15/8218)}
- C12N 15/8218: {Antisense, co-suppression, viral induced gene silencing [VIGS], post-transcriptional induced gene silencing [PTGS]}
- C12N 15/8286: {for insect resistance}
- C12N 2310/14: interfering nucleic acids [NA]
- Y02A 40/146: Genetically Modified [GMO] plants, e.g. transgenic plants
- 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)
- C12N: MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA (microbiological testing media C12Q1/00)
- Y02A: TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE

## Abstract

Disclosed are double stranded RNA molecules that are toxic to hemipteran insects. In particular, interfering RNA molecules capable of interfering with pest target genes and that are toxic to the target pest are provided. Further, methods of making and using the interfering RNA, for example in transgenic plants or as the active ingredient in a composition, to confer protection from insect damage are disclosed.

## Claims

### Claim 1

An interfering ribonucleic acid (RNA) molecule wherein the RNA comprises at least one dsRNA wherein the dsRNA is a region of double-stranded RNA comprising annealed complementary strands, one strand of which comprises a sequence of at least 30 contiguous nucleotides which is complementary to a target nucleotide sequence within a stink bug target gene, and (i) comprises at least a 30 contiguous nucleotide fragment of a sequence according to SEQ ID NO: 242, 296, or the complement thereof; or (ii) comprises at least a 30 contiguous nucleotide fragment of a nucleotide sequence encoding an amino acid sequence encoded by a sequence according to SEQ ID NO: 242, 296, or the complement thereof, wherein the interfering RNA molecule has insecticidal activity on a hemipteran plant pest.

### Claim 2 (depends on 1)

The interfering RNA molecule of claim 1, wherein said hemipteran plant pest is a stink bug.

### Claim 3 (depends on 1)

The interfering RNA molecule of claim 1, wherein the RNA comprises at least two dsRNAs, wherein each dsRNA comprises a sequence of nucleotides which is at least partially complementary to a target nucleotide sequence within the target gene.

### Claim 4 (depends on 1)

The interfering RNA molecule of claim 1, wherein the interfering RNA molecule comprises SEQ ID NO: 242, 296, or the complement thereof.

### Claim 5 (depends on 1)

The interfering RNA molecule of claim 1, wherein the stink bug is selected from the group consisting of Euschistus heros, Nezara viridula, Piezodorus guildinii, Halyomorpha halys, Chinavia hiare, Euschistus servus, Dichelops melacanthus, Dichelops furcatus, Edessa meditabunda, Thyanta perditor, Chinavia marginatum, Horcias nobilellus, Taedia stigmosa, Dysdercus peruvianus, Neomegalotomus parvus, Leptoglossus zonatus, and Niesthrea sidea.

### Claim 6 (depends on 1)

A nucleic acid construct comprising the interfering RNA molecule of claim 1.

### Claim 7 (depends on 1)

A nucleic acid molecule encoding the interfering RNA molecule of claim 1.

### Claim 8 (depends on 1)

A recombinant vector comprising a regulatory sequence operably linked to a nucleotide sequence that encodes the interfering RNA molecule of claim 1.

### Claim 9 (depends on 1)

A composition comprising two or more of the interfering RNA molecules of claim 1.

### Claim 10 (depends on 1)

An insecticidal composition for inhibiting the expression of a stink bug target gene, comprising the interfering RNA of claim 1 and an agriculturally acceptable earner.

### Claim 11 (depends on 1)

A transgenic plant, or part thereof, comprising the interfering RNA molecule, the nucleic acid molecule, the nucleic acid construct, and/or the composition of claim 1, wherein the transgenic plant has enhanced resistance to a stink bug as compared to a control plant.

### Claim 12 (depends on 11)

A transgenic seed of a transgenic plant of claim 11.

### Claim 13 (depends on 11)

A commodity product derived from the transgenic plant, or part thereof, of claim 11.

### Claim 14 (depends on 1)

A method of controlling a stink bug comprising contacting the stink bug with a nucleic acid molecule that is or is capable of producing an interfering RNA molecule of claim 1 for inhibiting expression of a target gene in the stink bug thereby controlling the stink bug.