Ras opposite (ROP) and related nucleic acid molecules that confer resistance to coleopteran and/or hemipteran pests

Patent metadata

Field	Value
Publication number	US-10647994-B2
Application number	US-201414577811-A
Country	US
Kind code	B2
Filing date	Dec 19, 2014
Priority date	Dec 20, 2013
Publication date	May 12, 2020
Grant date	May 12, 2020

Abstract

This disclosure concerns nucleic acid molecules and methods of use thereof for control of coleopteran and/or hemipteran pests through RNA interference-mediated inhibition of target coding and transcribed non-coding sequences in coleopteran and/or hemipteran pests. The disclosure also concerns methods for making that express nucleic acid molecules useful for the control of coleopteran and/or hemipteran pests, and the plant cells and plants obtained thereby.

First claim

What is claimed is:

1. A double-stranded ribonucleic acid (dsRNA) molecule comprising a first polyribonucleotide consisting of at least 23 contiguous nucleotides of the polyribonucleotide encoded by a polynucleotide selected from the group consisting of SEQ ID NO:1, SEQ ID NO:115, SEQ ID NO:120, SEQ NO:122, SEQ ID NO:124, SEQ ID NO:126, SEQ ID NO:131, and SEQ ID NO:133,
wherein the first polyribonucleotide is hybridized in the dsRNA molecule to a second polyribonucleotide that is the complement or reverse complement of the first polyribonucleotide, and
wherein delivery of the dsRNA molecule inhibits the expression of a target gene in a coleopteran or hemipteran insect selected from the group consisting of Diabrotica virgifera, Euschistus heros , and Meligethes aeneus.

2. The dsRNA molecule of claim 1 , wherein the second polyribonucleotide is the reverse complement of the first polyribonucleotide, and wherein the dsRNA molecule comprises a third polyribonucleotide separating the first and second polyribonucleotides in a single transcript.

3. The dsRNA molecule of claim 2 , where the dsRNA has a hairpin structure comprising a stem and a loop, wherein the hybridized first and second polyribonucleotides form the stein and the third polyribonucleotide forms the loop in the dsRNA molecule.

4. The dsRNA molecule of claim 1 , wherein the dsRNA is a siRNA, shRNA, miRNA, or hpRNA.

5. The dsRNA molecule of claim 1 , wherein the RNA molecule comprises a nucleic acid analogue.

6. The dsRNA molecule of claim 1 , wherein the first polyribonucleotide is encoded by a polynucleotide selected from the group of SEQ ID NO:3, SEQ ID NO:4, SEQ ID NO:114, SEQ ID NO:119, and SEQ ID NO:128.

7. A plant cell comprising the dsRNA molecule of claim 1 .

8. The plant cell of claim 7 , wherein the plant is selected from the group consisting of corn, maize, soybean, and plants of the family Poaceae.

9. A nucleic acid molecule encoding the dsRNA molecule of claim 1 , comprising a polynucleotide encoding the first polyribonucleotide, wherein the polynucleotide is operably linked to a heterologous promoter.

10. The nucleic acid molecule of claim 9 , wherein the molecule is a vector.

11. A method of controlling a population of coleopteran or hemipteran pests selected from the group consisting of Diabrotica virgifera, Euschistus heros , and Meligethes aeneus , the method comprising:
feeding the dsRNA molecule of claim 1 to insects of the population.

12. A food source for an insect pest comprising the dsRNA molecule of claim 1 .

13. The method according to claim 11 wherein feeding the dsRNA molecule to insects of the population comprises feeding the insects with an artificial diet food source comprising the dsRNA molecule.

14. The method according to claim 11 , wherein feeding the dsRNA molecule to insects of the population comprises spraying a plant infested with the insects with a formulation comprising the dsRNA molecule.

15. The dsRNA molecule of claim 2 , wherein the dsRNA molecule is encoded by a polynucleotide selected from the group consisting of SEQ ID NOs:13-15.

16. A nucleic acid molecule encoding the dsRNA molecule of claim 2 , comprising a polynucleotide encoding the dsRNA, wherein the polynucleotide is operably linked to a promoter.

17. The nucleic acid molecule of claim 16 , wherein the molecule is a plant transformation vector.

18. The nucleic acid molecule of claim 16 , wherein the polynucleotide is selected from the group consisting of SEQ ID NOs:13-15.

19. A plant cell comprising the nucleic acid molecule of claim 16 .

20. The plant cell of claim 19 , wherein the plant is selected from the group consisting of corn, maize, soybean, and plants of the family Poaceae.

21. The plant cell of claim 19 , wherein the nucleic acid molecule is integrated into the genome of the plant cell.

22. A plant comprising the plant cell of claim 21 .

23. A method of controlling a population of coleopteran or hemipteran pests selected from the group consisting of Diabrotica virgifera, Euschistus heros , and Meligethes aeneus , the method comprising:
feeding the plant cell of claim 19 to insects of the population.

24. The method according to claim 23 , wherein the plant cell is present in a plant material or whole plant.

25. The method according to claim 23 , wherein the plant cell is of a plant selected from the group consisting of corn, maize, soybean, and plants of the family Poaceae.

Assignees

• Dow Agrosciences Llc
• Fraunhofer Ges Forschung

Inventors

• Narva Kenneth E
• Li Huarong
• Geng Chaoxian
• Arora Kanika
• Veeramani Balaji
• Gandra Premchand
• Worden Sarah
• Vilcinskas Andreas
• Knorr Eileen
• Fishilevich Elane
• Rangasamy Murugesan
• Frey Meghan

Classifications

• C12N2310/14

  interfering nucleic acids [NA] · CPC title

• C12N15/8286 Primary

  for insect resistance · CPC title

• C12N15/8218

  Antisense, co-suppression, viral induced gene silencing [VIGS], post-transcriptional induced gene silencing [PTGS] · CPC title

• C12N2310/531

  Stem-loop; Hairpin · CPC title

• C12N15/113 Primary

  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 title