Compositions and methods for controlling arthropod parasite and pest infestations

• US 9,540,642 B2
• Filed: 11/04/2014
• Issued: 01/10/2017
• Est. Priority Date: 11/04/2013
• Status: Active Grant

• Current Assignee: GreenLight Biosciences, Inc., The United States of America As Represented By The Secretary of Agriculture
• Original Assignee: Beeologics Inc. (Bayer AG), Monsanto Technology LLC (Bayer AG), United States Department Of Agriculture
• Inventors: Inberg, Alex, Kapoor, Mahak, Evans, Jay
• Primary Examiner(s): VIVLEMORE, TRACY ANN
• Application Number:
  US14/532,596
• Publication Number:
  US 20150159156A1
• Time in Patent Office: 798 Days
• Field of Search: None
• US Class Current: 1/1
• CPC Class Codes: A01K 51/00 Appliances for treating beehives or parts thereof, e.g. for cleaning or disinfecting
• CPC Class Codes: A01N 37/46 N-acyl derivatives
• CPC Class Codes: A01N 63/14 Insects
• CPC Class Codes: A01N 63/16 Arachnids
• CPC Class Codes: A01N 63/60 Isolated nucleic acids
• CPC Class Codes: A61K 31/713 Double-stranded nucleic acids or oligonucleotides
• 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 2310/14 interfering N.A.
• CPC Class Codes: C12N 2310/531 Stem-loop; Hairpin

Abstract

This application provides and discloses anti-parasitic, anti-pest or insecticidal nucleic acid molecules and their calmodulin target genes for the control of arthropod parasites and pests. This application further provides methods and compositions for the control and treatment of parasites and pests in Apis mellifera (honey bee) hives.

Claims

1. A bee-ingestible, bee-absorbable, mite-ingestible, or mite-absorbable composition comprising an excipient and a nucleic acid molecule having a sequence that is at least 96% identical or complementary to at least 21 contiguous nucleotides of SEQ ID NO:

3.

2. The composition of claim 1, wherein said nucleic acid sequence is a dsRNA.

3. The composition of claim 1, wherein said excipient is selected from the group consisting of protein, pollen, carbohydrate, polymer, liquid solvent, sugar syrup, sugar solid, and semi-solid feed.

4. The composition of claim 2, wherein said dsRNA is an siRNA.

5. The composition of claim 1, wherein said nucleic acid sequence comprises at least 23 contiguous nucleotides of SEQ ID NO:

3.

6. The composition of claim 2, wherein said dsRNA sequence comprises at least 23 contiguous nucleotides of SEQ ID NO:

3.

10. A method of providing a composition to a honeybee, comprising providing the bee an effective amount of the composition of claim 1, whereby the nucleic acid is present in honeybee tissue.

11. The method of claim 10, wherein said calmodulin gene sequence is a Varroa destructor calmodulin gene sequence.

12. The method of claim 10, wherein said honeybee is a forager or a hive bee.

13. The method of claim 11, wherein said honeybee is a bee of a colony and said providing reduces the susceptibility of said bee colony to Varroa destructor, reduces the parasitation of said bee colony by Varroa destructor, or reduce the parasite load of said bee colony by Varroa destructor.

14. The method of claim 11, wherein said providing reduces the susceptibility of said honeybee to Varroa destructor, reduces the parasitation of said honeybee by Varroa destructor, or reduce the parasite load of said honeybee by Varroa destructor.

15. The composition of claim 1, further comprising one or more nucleic acid molecules having a second nucleic acid sequence that is essentially identical or essentially complementary to a different region of a calmodulin gene sequence.

7. A nucleic acid construct comprising a nucleic acid that encodes a sequence that is at least 95% identical or complementary to SEQ ID NO:

3 operably linked to a promoter sequence functional in a host cell and capable of producing a dsRNA when introduced into said host cell.

8. The nucleic acid construct of claim 7, further comprising at least one regulatory element selected from the group consisting of translation leader sequences, introns, enhancers, stem-loop structures, repressor binding sequences, termination sequences, pausing sequences, and polyadenylation recognition sequences.

9. The nucleic acid construct of claim 7, wherein said host cell is selected from the group consisting of a bacteria cell and a yeast cell.