dsRNA delivery composition and methods of use
View Patent ↗The present invention provides for compositions comprising a cucurbitacin bait for stimulating insect feeding and a dsRNA—such as for eliciting RNAi mediated gene silencing in an insect. The present invention also provides for methods of delivering a dsRNA molecule to an insect, inhibiting protein expression in an insect, and killing an insect by providing a dsRNA molecule.
1. A composition for delivering a dsRNA molecule to an insect, the composition comprising a cucurbitacin bait and at least one DvvEST3 dsRNA molecule, wherein the DvvEST3 dsRNA molecule elicits an RNAi response in the insect.
2. The composition of claim 1 wherein the composition comprises at least two dsRNA molecules of different sequences.
3. The composition of claim 1 wherein the composition further comprises a vATPase subunit A-dsRNA molecule that elicits an RNAi response in the insect.
4. The composition of claim 3 wherein at least one nucleic acid sequence selected from the group consisting of SEQ ID NO:6 and SEQ ID NO:7 is used as a template for synthesis of the vATPase subunit A-dsRNA molecule.
5. The composition of claim 1 wherein at least one nucleic acid sequence selected from the group consisting of SEQ ID NO:8 and SEQ ID NO: 9 is used as a template for synthesis of the DvvEST3 dsRNA molecule.
6. The composition of claim 1 wherein the composition comprises from about 5% to about 50% of the cucurbitacin bait.
7. The composition of claim 6 wherein the composition comprises from about 5% to about 25% of the cucurbitacin bait.
8. The composition of claim 6 wherein the composition comprises at least about 4% agar.
9. A method of delivering a dsRNA molecule to an insect, the method comprising providing an insect with a composition that comprises a cucurbitacin bait and at least one DvvEST3 dsRNA molecule to feed upon, wherein the DvvEST3 dsRNA molecule elicits an RNAi response in the insect and wherein the insect is selected from the group consisting of western corn rootworm, Diabrotica balteata LeConte, banded cucumber beetle, Diabrotica barberi Smith and Lawrence, northern corn rootworm, Diabrotica undecimpunctata howardi Barber, southern corn rootworm, and Diabrotica speciosa.
10. The method of claim 9 wherein the insect is a western corn rootworm.
11. The method of claim 9 wherein the composition comprises at least two dsRNA molecules of different sequences that elicit an RNAi response in the insect.
12. The method of claim 9 wherein the composition comprises about 5% to about 50% of the cucurbitacin bait.
13. The method of claim 9 wherein the dsRNA molecule increases mortality of the insect.
14. The method of claim 9 wherein the composition provided to the insect further comprises at least one vATPase subunit A-dsRNA molecule to feed upon that elicits a dsRNA response in the insect.
15. The method of claim 14 wherein at least one nucleic acid sequence selected from the group consisting of SEQ ID NO:6 and SEQ ID NO:7 is used as a template for synthesis of the vATPase subunit A-dsRNA molecule.
16. The method of claim 9 wherein at least one nucleic acid sequence selected from the group consisting of SEQ ID NO:8 and SEQ ID NO:9 is used as a template for synthesis of the DvvEST3 dsRNA molecule.
17. A method of producing a composition for delivering a dsRNA molecule to an insect, the method comprising combining a cucurbitacin bait and at least one DvvEST3 dsRNA molecule, wherein the DvvEST3 dsRNA molecule elicits an RNAi response in an insect.
18. The method of claim 17 wherein the composition comprises at least two dsRNA molecules of different sequences that elicit an RNAi response in the insect.
19. The method of claim 17 wherein at least one nucleic acid sequence selected from the group consisting of SEQ ID NO:8 and SEQ ID NO:9 is used as a template for synthesis of the DvvEST3 dsRNA molecule.
20. The method of claim 17 wherein the method further comprises combining a vATPase subunit A-dsRNA molecule that elicits an RNAi response in the insect with the cucurbitacin bait and at least one DvvEST3 dsRNA molecule.
21. The method of claim 20 wherein at least one nucleic acid sequence selected from the group consisting of SEQ ID NO:6 and SEQ ID NO:7 is used as a template for synthesis of the vATPase subunit A-dsRNA molecule.
22. The method of claim 17 wherein the composition produced comprises from about 5% to about 50% of the cucurbitacin bait.