Antisense oligonucleotide and double stranded RNAS for control of hemipteran and lepidopteran pests

Patent metadata

Field	Value
Publication number	US-11795458-B2
Application number	US-202117554026-A
Country	US
Kind code	B2
Filing date	Dec 17, 2021
Priority date	Dec 17, 2020
Publication date	Oct 24, 2023
Grant date	Oct 24, 2023

Abstract

The present disclosure provides oligonucleotides and double-stranded RNAs (dsRNAs) targeting Hemipteran and Lepidopteran insect pests. Bantam-sequence targets are detailed and shown to be effective targets for RNAi induction utilizing multiple dsRNAs, antisense oligonucleotides, and modified dsRNAs (e.g., 2′-O-methylated and dsRNAs containing non-canonical nucleosides).

First claim

The embodiment of the invention in which exclusive property or privilege is claimed is defined as follows:

1. A double-stranded ribonucleic acid (dsRNA) comprising a first strand comprising a sequence with at least 95% identity to any one of SEQ ID NO:5, SEQ ID NO: 6; or SEQ ID NO: 7 and a second strand complementary to the first strand.

2. The dsRNA of claim 1 , wherein the first strand has at least 99% or 100% sequence identity to any one of SEQ ID NO:5, SEQ ID NO: 6; or SEQ ID NO: 7.

3. A double-stranded ribonucleic acid (dsRNA) comprising a first strand comprising SEQ ID NO:3 and a second strand complementary to the first strand.

4. A double-stranded ribonucleic acid (dsRNA) comprising a first strand comprising SEQ ID NO:4 and a second strand complementary to the first strand.

5. The dsRNA of claim 1 , wherein the dsRNA is capable of inducing ribonucleic acid interference (RNAi) when ingested by a hemipteran or lepidopteran insect.

6. The dsRNA of claim 5 , wherein the insect is S. frugiperda or D. citri.

7. The dsRNA of claim 1 , wherein the dsRNA comprises at least one base comprising a 2′-O-methyl moiety.

8. The dsRNA of claim 1 , wherein the dsRNA comprises at least one non-canonical nucleotide.

9. The dsRNA of claim 8 , wherein the non-canonical nucleotide is 2′-fluorine-dCTP or 2′-fluorine-dUTP.

10. The dsRNA of claim 9 , wherein each cytosine comprises 2′-fluorine-dCTP and each uracil comprises 2′-fluorine-dUTP.

11. A DNA molecule comprising a promoter functional in a host cell and a DNA encoding a dsRNA comprising a first strand and a second strand, wherein the first strand comprises a sense region with at least 95% sequence identity to any one of SEQ ID NO:5, SEQ ID NO: 6; or SEQ ID NO: 7 and a second strand complementary to the first strand.

12. A method of inducing RNAi in a hemipteran or lepidopteran insect, comprising contacting the dsRNA of claim 1 with a hemipteran or lepidopteran insect such that the insect ingests the dsRNA, thereby inducing RNAi in the insect.

13. The method of claim 12 , wherein the insect is S. frugiperda or D. citri.

14. A method of inducing RNAi in a hemipteran or lepidopteran insect, comprising contacting a dsRNA with a hemipteran or lepidopteran insect such that the insect ingests the dsRNA, thereby inducing RNAi in the insect, wherein the first strand of the dsRNA comprises SEQ ID NO: 3, SEQ ID NO: 4, SEQ ID NO: 5, or SEQ ID NO: 6.

15. The method of claim 14 , wherein the first strand of the dsRNA comprises SEQ ID NO: 4.

16. The method of claim 14 , wherein the first strand of the dsRNA comprises SEQ ID NO: 5.

17. The method of claim 14 , wherein the first strand of the dsRNA comprises SEQ ID NO: 6.

18. The method of claim 14 , wherein the first strand of the dsRNA comprises SEQ ID NO: 3.

Assignees

• Us Agriculture

Inventors

• Hunter Wayne B
• Lopez Salvador P

Classifications

• 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

• A01N63/60

  Isolated nucleic acids · CPC title

• A01P7/04

  Insecticides · CPC title

• C12N2310/11

  Antisense · CPC title

• C12N2310/321

  2'-O-R Modification · CPC title