Compositions for conferring tolerance to viral disease in social insects, and the use thereof

• US 20090118214A1
• Filed: 08/20/2008
• Published: 05/07/2009
• Est. Priority Date: 11/07/2007
• Status: Active Grant

• Current Assignee: GreenLight Biosciences, Inc.
• Original Assignee: Beeologics LLC (Bayer AG)
• Inventors: Yarden, Gal, Paldi, Nitzan
• Granted Patent: US 8,097,712 B2
• Time in Patent Office: Days
• US Class Current: 514/44.R
• CPC Class Codes: A61K 31/713 Double-stranded nucleic acids or oligonucleotides
• CPC Class Codes: A61P 31/00 Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
• CPC Class Codes: A61P 31/12 Antivirals
• 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 15/1131 against viruses
• CPC Class Codes: C12N 2310/14 interfering N.A.
• CPC Class Codes: C12N 2770/22022 New viral proteins or individual genes, new structural or functional aspects of known viral proteins or genes

Abstract

Compositions and methods for reducing susceptibility to infectious disease in bees using RNA interference technology, and more particularly, prevention and treatment of viral infections in honeybees such as Israel acute paralysis virus (IAPV) by feeding of pathogen-specific dsRNA. Further, multiple-pathogen specific dsRNA is disclosed.

Claims

1. An isolated nucleic acid agent comprising a nucleic acid sequence downregulating expression of a gene product of a bee pathogen.

2. The isolated nucleic acid of claim 1, wherein said gene product is an mRNA encoding a bee pathogen specific polypeptide.

3. The isolated nucleic acid agent of claim 1, wherein the agent is selected from the group consisting of a dsRNA, an antisense RNA and a ribozyme.

4. The isolated nucleic acid agent of claim 3, wherein said dsRNA is selected from the group consisting of siRNA, shRNA and miRNA.

5. A nucleic acid construct comprising a nucleic acid sequence encoding the isolated nucleic acid agent of claim 1.

6. The isolated nucleic acid agent of claim 1, wherein said nucleic acid sequence is greater than 15 base pairs in length.

7. The isolated nucleic acid agent of claim 6, wherein said nucleic acid sequence is 19 to 25 base pairs in length.

8. The isolated nucleic acid agent of claim 6, wherein said nucleic acid sequence is greater than 30 base pairs in length.

9. The isolated nucleic acid agent of claim 1, wherein said bee pathogen is selected from the group consisting of a virus, a bacteria, a parasitic protozoan, a fungus and a nematode.

10. The isolated nucleic acid agent of claim 1, wherein said bee pathogen is a virus.

11. The isolated nucleic acid of claim 10, wherein said virus is Israel Acute Paralysis Virus.

12. The isolated nucleic acid of claim 10, wherein said virus is Kashmir Paralysis Virus.

13. The isolated nucleic acid of claim 10, wherein said virus is Israel Acute Paralysis Virus and said polypeptide of said virus is selected from the group consisting of IAPV polymerase polyprotein (SEQ ID NO. 51) and IAPV structural polyprotein (SEQ ID NO:

52).

14. The isolated nucleic acid of claim 1, wherein said viral pathogen is Israel Acute Paralysis Virus and said nucleic acid sequence is as set forth in SEQ ID NOs:

24, 33 and 34.

15. The isolated nucleic acid of claim 1, wherein said viral pathogen is Israel Acute Paralysis Virus and said nucleic acid sequence is as set forth in SEQ ID NOs:

54-9532.

16. The isolated nucleic acid of claim 1, wherein said virus is Israel Acute Paralysis Virus, and said nucleic acid sequence is a viral nucleic acid sequence integrated into a honeybee genome following Israel Acute Paralysis Virus infection.

17. A bee-ingestible composition comprising the nucleic acid agent of claim 1.

18. The bee-ingestible composition of claim 17, wherein said composition is in solid form.

19. The bee-ingestible composition of claim 17, wherein said composition is in liquid form.

20. The bee-ingestible composition of claim 17, wherein said composition comprises protein.

21. The bee-ingestible composition of claim 20, wherein said protein is in the form of pollen and/or soy patties.

22. The bee-ingestible composition of claim 19, wherein said liquid is a sucrose solution.

23. The bee-ingestible composition of claim 19, wherein said liquid is a corn syrup solution.

24. The bee-ingestible composition of claim 19, wherein said liquid further comprises a carbohydrate or sugar supplement.

25. A method for reducing the susceptibility of a bee to a disease caused by a pathogen comprising feeding the bee an effective amount of the nucleic acid agent of claim 1, thereby reducing the susceptibility of said bee to said pathogen.

26. The method of claim 25, wherein said bee is a honeybee.

27. The method of claim 25, wherein said honeybee is a forager.

28. The method of claim 25, wherein said honeybee is a hive bee.

29. The method of claim 25, wherein said honeybee is a bee of a colony, wherein said feeding reduces the susceptibility of said bee colony to said disease.

30. The method of claim 25, wherein said disease is Colony Collapse Disorder.

31. The method of claim 25, wherein said bee pathogen is Israel Acute Paralysis Virus.

32. The method of claim 31, wherein said nucleic acid agent or nucleic acid construct comprises a nucleic acid sequence selected from the group consisting of SEQ ID NOs:

54-9532.

33. The method of claim 31, wherein said nucleic acid agent or nucleic acid construct comprises a nucleic acid sequence as set forth in SEQ ID NOs:

24, 33 and 34.

34. The method of claim 25, wherein said feeding comprises providing a liquid bee-ingestible composition.

35. The methods of claim 25, wherein said feeding comprises providing a solid bee-ingestible composition.

36. A method for reducing the susceptibility of a bee to a disease caused by a pathogen comprising feeding the bee an effective amount of the nucleic acid construct of claim 5, thereby reducing the susceptibility of said bee to said pathogen.

37. The method of claim 36, wherein said nucleic acid construct comprises a nucleic acid sequence selected from the group consisting of SEQ ID NOs:

54-9532.

38. The method of claim 36, wherein said nucleic acid construct comprises a nucleic acid sequence as set forth in SEQ ID NOs:

24, 33 and 34.

39. A method of reducing the susceptibility of honeybees to Colony Collapse Disorder (CCD), the method comprising feeding to the honeybee hive an effective amount of double stranded ribonucleic nucleic acid (RNA), said double stranded RNA comprising a sequence complementary to at least 21 nucleotides of an Israel Acute Paralysis Virus-specific mRNA and capable of inducing degradation of said virus-specific mRNA.

40. An isolated nucleic acid agent comprising a nucleic acid sequence complementary to at least 21 nucleotides of a bee pathogen specific RNA and capable of inducing degradation of said bee pathogen specific RNA.

41. An isolated nucleic acid agent comprising at least two nucleic acid sequences downregulating expression of at least one gene product of at least one bee pathogen.

42. The isolated nucleic acid of claim 41, wherein said at least one bee pathogen is selected from the group consisting of a virus, a bacteria, a parasitic protozoan, a fungus and a nematode.

43. The isolated nucleic acid of claim 41, wherein said at least one bee pathogen comprises at least two bee viruses.

44. The isolated nucleic acid of claim 41, wherein said nucleic acid sequences correspond to conserved sequences in said at least two bee pathogens.

45. The isolated nucleic acid of claim 41, wherein said isolated nucleic acid sequence is as set forth in SEQ ID NO. 24.