Targeted control of pests and pathogens by plant delivery of 2′F-ANA-oligonucleotides

Patent metadata

Field	Value
Publication number	US-11946058-B2
Application number	US-202217568884-A
Country	US
Kind code	B2
Filing date	Jan 5, 2022
Priority date	Jul 6, 2018
Publication date	Apr 2, 2024
Grant date	Apr 2, 2024

Abstract

Herein is disclosed synthetic oligonucleotides comprising 2′F-ANA nucleosides that can be utilized to control plant-chewing and phloem-feeding insects, bacteria present in such insects, and bacteria present in plants. The novel approaches and materials provided herein allow for reduction of pesticide and antibiotic use without the need to create genetically modified plants.

First claim

What is claimed is:

1. A method of inducing RNA silencing in a plant-chewing or phloem-feeding insect comprising the steps of:
a. providing an oligonucleotide comprising at least one 2′F-ANA-modified nucleotide and at least one 2′-deoxyribonucleotide, and wherein the 2′F-ANA-modified nucleotides are positioned according to any of Formula 2-16, and having the sequence of SEQ ID NO: 39-46, SEQ ID NO: 48-69 and SEQ ID NO: 71-76 to the insect in a manner whereby the insect to ingests the oligonucleotide, thereby inducing RNA silencing.

2. The method of claim 1 , wherein the insect ingests the oligonucleotide by consuming plant material containing the oligonucleotide.

3. The method of claim 1 , wherein the insect is Diaphorina citri or Diaprepes abbreviatus.

4. The method of claim 1 , wherein the plant is a citrus plant or a potato plant.

5. The method of claim 4 , wherein plant is a citrus plant selected from the group consisting of orange, lemon, clementine, lime, grapefruit, pomelo, citron, mandarin, and tangelo.

6. The method of claim 1 , comprising the additional step of applying the oligonucleotide to a plant.

7. The method of claim 6 , wherein the oligonucleotide is applied to a plant by root soak, injection or foliar spray.

8. The method of claim 1 , comprising the additional step of applying the oligonucleotide to an artificial diet, sugar solution or bait material.

9. A method of controlling a plant-chewing or phloem-feeding insect, comprising the steps of:
a. providing an oligonucleotide comprising at least one 2′F-ANA-modified nucleotide and at least one 2′-deoxyribonucleotide, and wherein the 2′F-ANA-modified nucleotides are positioned according to any of Formula 2-16, and having a sequence selected from the group consisting of SEQ ID NO: 39-46, SEQ ID NO: 48-69 and SEQ ID NO: 71-76 in a manner whereby the insect to ingests the oligonucleotide, thereby inducing RNA silencing and a detrimental effect to the insect.

10. The method of claim 9 , wherein the detrimental effect is increased mortality compared to insects not exposed to the oligonucleotide.

11. The method of claim 9 , wherein the insect is Diaphorina citri or Diaprepes abbreviatus.

12. The method of claim 9 , wherein the providing step comprises applying the oligonucleotide to a plant.

13. The method of claim 9 , wherein the providing step comprises applying the oligonucleotide to an artificial diet, sugar solution or bait material.

14. A method of inducing RNA silencing in a Diaphorina citri or Diaprepes abbreviatus insect comprising the steps of:
a. contacting D. cirri or D. abbreviatus with an oligonucleotide comprising at least one 2′F-ANA-modified nucleotide and at least one 2′-deoxyribonucleotide in an amount sufficient to induce RNA silencing, wherein the 2′F-ANA-modified nucleotides are positioned according to any of Formulas 2-16.

15. The method of claim 14 , wherein the oligonucleotide is selected from the group consisting of SEQ ID NO: 39-46, SEQ ID NO: 48-69 and SEQ ID NO: 71-76.

16. The method of claim 14 , wherein the insect is D. citri and the contacting comprises ingestion of phloem of a plant, wherein the phloem comprises the oligonucleotide.

17. The method of claim 14 , wherein the insect is D. abbreviatus and the contacting comprises consumption of plant tissue, wherein the plant tissue comprises the oligonucleotide.

Assignees

• Us Agriculture
• Aum Lifetech Inc

Inventors

• Hunter Wayne B
• Aishwarya Veenu

Classifications

• C12N15/8218 Primary

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

• C12N2310/11

  Antisense · CPC title

• C12N2310/323

  modified ring structure · CPC title

• C12N2310/346

  having a combination of backbone and sugar modifications · CPC title