Down-regulating gene expression in insect pests

• US 9,914,925 B2
• Filed: 11/03/2015
• Issued: 03/13/2018
• Est. Priority Date: 04/20/2011
• Status: Expired due to Fees

• Current Assignee: deVGen NV (China National Chemical Corporation)
• Original Assignee: deVGen NV (China National Chemical Corporation)
• Inventors: Beghyn, Myriam, Bogaert, Thierry Andre Olivier Eddy, Feldmann, Pascale, Raemaekers, Romaan
• Primary Examiner(s): DEVEAU ROSEN, JASON
• Application Number:
  US14/930,753
• Publication Number:
  US 20160060628A1
• Time in Patent Office: 861 Days
• Field of Search: None
• CPC Class Codes: A01N 57/16 containing heterocyclic radicals
• CPC Class Codes: A01N 63/60 Isolated nucleic acids
• CPC Class Codes: C07K 14/43563 from insects
• CPC Class Codes: 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
• CPC Class Codes: C12N 15/8218 Antisense, co-suppression, viral induced gene silencing [VIGS], post-transcriptional induced gene silencing [PTGS]
• CPC Class Codes: C12N 15/8286 for insect resistance
• CPC Class Codes: C12N 2310/14 interfering N.A.
• CPC Class Codes: Y02A 40/146 Genetically Modified [GMO] plants, e.g. transgenic plants

Abstract

The present invention relates to genetic control of infestation by insect pest species, particularly prevention and/or control of pest infestation of plants, using interfering ribonucleic acid (RNA) molecules. Compositions and combinations containing the interfering RNA molecules of the invention for use in topical applications, for example in the form of insecticides.

Claims

1. An interfering ribonucleic acid (RNA) that functions upon uptake by an insect pest species to down-regulate expression of a ribosomal protein S27 target gene in said insect pest, wherein the RNA comprises at least one silencing element, wherein the silencing element is a region of double-stranded RNA comprising annealed complementary strands, one strand of which comprises or consists of a sequence of nucleotides which is at least partially complementary to a target nucleotide sequence within the target gene, and wherein the interfering RNA is encoded by:

(i) a nucleotide sequence comprising any of SEQ ID NOs 15, 16, or the complement thereof, or a nucleotide sequence that is at least 95% identical to any of SEQ ID NOs 15, 16, or the complement thereof,

(ii) a nucleotide sequence comprising a fragment of at least 21 contiguous nucleotides of any of SEQ ID NOs 15, 16, the complement thereof,

(iii) a nucleotide sequence comprising a fragment of at least 21 contiguous nucleotides of any of SEQ ID NOs 15, 16, or the complement thereof, and wherein said nucleotide sequence of said fragment is at least 95% identical to said corresponding fragment of any of SEQ ID NOs 15, 16, or the complement thereof, or

(iv) a nucleotide sequence comprising a fragment of at least 21 contiguous nucleotides of a nucleotide sequence encoding an amino acid sequence encoded by SEQ ID NO;

15, 16, the complement thereof.

2. The interfering RNA of claim 1 wherein the silencing element comprises or consists of a sequence of at least 21 contiguous nucleotides which is complementary or at least partially complementary to a target nucleotide sequence within a ribosomal protein S27 target gene.

3. The interfering RNA of claim 1 wherein the RNA comprises at least two silencing elements, wherein each silencing element comprises or consists of a sequence of nucleotides which is at least partially complementary to a target nucleotide sequence within the ribosomal protein S27 target gene.

4. The interfering RNA of claim 3 wherein each of the silencing elements comprises or consists of a different sequence of nucleotides which is complementary to a different target nucleotide sequence within the ribosomal protein S27 target gene.

5. The interfering RNA of claim 4 wherein the different ribosomal protein S27 target nucleotide sequences originate from a single ribosomal protein S27 target gene or from different target genes.

6. The interfering RNA of claim 5 wherein the different target genes originate from the same insect pest species or different insect pest species.

7. The interfering RNA of claim 1 wherein the insect pest species is a plant pest.

8. The interfering RNA of claim 1 wherein the plant pest is an insect pest species selected from the insect species belonging to the orders Coleoptera, Hemiptera, Lepidoptera, Diptera, Dichyoptera, Orthoptera, and Siphonaptera.

9. The interfering RNA of claim 8 wherein the insect plant pest is selected from the group consisting of Leptinotarsa spp., Nilaparvata spp., Lygus spp., Myzus spp., and Diabrotica spp.

10. The interfering RNA of claim 1 wherein down-regulating expression of the ribosomal protein S27 target gene causes decreased growth, development, reproduction, or survival of the pest as compared with a pest species exposed to an interfering ribonucleic acid targeting a non-essential gene or an interfering ribonucleic acid that does not down-regulate any genes within the pest species.

11. A polynucleotide comprising a sequence of nucleotides encoding the interfering RNA of claim 1.

12. The polynucleotide of claim 11 which is comprised in a DNA construct.

13. The DNA construct of claim 12 which is an expression construct, wherein the polynucleotide sequence encoding the interfering RNA is operably linked to at least one regulatory sequence capable of driving expression of the polynucleotide sequence.

14. A host cell comprising the interfering RNA of claim 1.

15. The host cell of claim 14 wherein the host cell is a prokaryotic or a eukaryotic cell.

16. The host cell of claim 15 wherein the host cell is a bacterial cell.

17. A composition for preventing and/or controlling insect pest infestation comprising the interfering ribonucleic acid (RNA) of claim 1 and at least one suitable carrier, excipient or diluent.

18. The composition of claim 17 wherein the interfering RNA is encoded by a polynucleotide comprising a sequence of nucleotides encoding the interfering RNA which is comprised in a DNA construct, which optionally is an expression construct wherein the polynucleotide sequence encoding the interfering RNA is operably linked to at least one regulatory sequence capable of driving expression of the polynucleotide sequence.

19. The composition of claim 17, wherein the interfering RNA is produced by a host cell capable of expressing the interfering RNA.

20. The composition of claim 19 wherein the host cell is a bacterial cell.

21. The composition of claim 17 wherein the composition is in a form suitable for ingestion by an insect.

22. The composition of claim 17 wherein the composition is in solid, liquid or gel form.

23. The composition of claim 17 wherein the composition is formulated as an insecticidal spray.

24. The composition of claim 23 wherein the spray is a pressurized/aerosolized spray or a pump spray.

25. The composition of claim 17 wherein the composition further comprises at least one pesticidal agent selected from the group consisting of a chemical insecticide, a patatin, a Bacillus thuringiensis insecticidal protein, a Xenorhabdus insecticidal protein, a Photorhabdus insecticidal protein, a Bacillus laterosporus insecticidal protein, and a Bacillus sphaericus insecticidal protein.

26. The composition of claim 25 wherein said Bacillus thuringiensis insecticidal protein is selected from the group consisting of a Cry1, a Cry3, a TIC851, a CryET170, a Cry22, a TIC901, a TIC201, a TIC407, a TIC417, a binary insecticidal protein CryET80 and CryET76, a binary insecticidal protein TIC100 and TIC101, a combination of an insecticidal protein ET29 or ET37 with an insecticidal protein TIC810 or TIC812, and a binary insecticidal protein PS149B1.

27. A housing or trap for an insect pest which contains the composition as defined in claim 17.

28. A combination for preventing and/or controlling pest infestation comprising the composition of claim 17 and at least one other active agent.

29. The combination of claim 28 wherein the combination is for preventing and/or controlling pest infestation of a plant and the other active agent is an agronomical agent.

30. The combination of claim 29 wherein the agronomical agent comprises a herbicide.

31. The combination of claim 29 wherein the agronomical agent comprises a pesticide.

32. The combination of claim 31 further comprising a second pesticide, wherein the second pesticide is selected from the group consisting of a chemical insecticide, a patatin, a Bacillus thuringiensis insecticidal protein, a Xenorhabdus insecticidal protein, a Photorhabdus insecticidal protein, a Bacillus laterosporus insecticidal protein, and a Bacillus sphaericus insecticidal protein.

33. The combination of claim 32 wherein said Bacillus thuringiensis insecticidal protein is selected from the group consisting of a Cry1, a Cry3, a TIC851, a CryET170, a Cry22, a TIC901, a TIC201, a TIC407, a TIC417, a binary insecticidal protein CryET80 and CryET76, a binary insecticidal protein TIC100 and TIC101, a combination of an insecticidal protein ET29 or ET37 with an insecticidal protein TIC810 or TIC812, and a binary insecticidal protein PS149B1.

34. A method for down-regulating expression of a target gene in an insect pest species in order to prevent and/or control pest infestation, comprising contacting said pest species with at least one interfering ribonucleic acid (RNA), wherein the interfering RNA functions upon uptake by the pest to down-regulate the expression of a target gene within said pest, wherein the interfering RNA is the interfering RNA of claim 1.

35. The method of claim 34 wherein down-regulation of expression of a target gene in an insect pest species is used to obtain at least 20% pest control or at least 20% pest mortality as compared to control insect pests contacted with an interfering ribonucleic acid (RNA) targeting a non-essential pest gene or a target gene not expressed in said pest.

36. The method of claim 34 wherein the method is used to prevent and/or control pest infestation of a plant.

37. The method of claim 36 wherein the plant is selected from the group consisting of cotton, potato, rice, canola, sunflower, sorghum, pearl millet, corn, strawberries, soy, alfalfa, tomato, eggplant, pepper and tobacco.

38. A kit comprising the interfering ribonucleic acid (RNA) of claim 1, the DNA construct of claim 12, or the composition of claim 17 for preventing and/or controlling insect pest infestation.