Invertebrate microRNAs

Patent metadata

Field	Value
Publication number	US-10570414-B2
Application number	US-201615348839-A
Country	US
Kind code	B2
Filing date	Nov 10, 2016
Priority date	Feb 20, 2007
Publication date	Feb 25, 2020
Grant date	Feb 25, 2020

Abstract

This invention provides plants having resistance to invertebrate pests. More specifically, this invention discloses a non-natural transgenic plant cell expressing at least one invertebrate miRNA in planta for suppression of a target gene of an invertebrate pest or of a symbiont associated with the invertebrate pest. Also provided are recombinant DNA constructs for expression of at least one invertebrate miRNA in planta, a non-natural transgenic plant containing the non-natural transgenic plant cell of this invention, a non-natural transgenic plant grown from the non-natural transgenic plant cell of this invention, and non-natural transgenic seed produced by the non-natural transgenic plants, as well as commodity products produced from a non-natural transgenic plant cell, plant, or seed of this invention. This invention further provides a method of suppressing at least one target gene of an invertebrate pest of a plant or of a symbiont associated with the invertebrate, including providing a plant including the non-natural transgenic plant cell of this invention, wherein the invertebrate is the invertebrate pest, the recombinant DNA is transcribed in the non-natural transgenic plant cell to the recombinant miRNA precursor, and when the invertebrate pest ingests the recombinant miRNA precursor, the at least one target gene is suppressed.

First claim

What is claimed is:

1. A method of suppressing at least one target gene of an invertebrate pest of a plant, comprising:
(a) providing in said plant a recombinant miRNA precursor comprising a single strand of RNA that folds into a secondary structure of a miRNA precursor identified from an invertebrate pest genome, wherein said miRNA precursor comprises at least one stem-loop capable of being processed into a mature miRNA capable of suppressing at least one target gene of said invertebrate pest; and
(b) providing said plant to said invertebrate pest, wherein ingestion of said mature miRNA by said invertebrate pest results in suppression of said at least one target gene.

2. The method of claim 1 , wherein said recombinant miRNA precursor comprises a miRNA selected from the group consisting of bta-miR-7 (SEQ ID NO: 70), dme-miR-9a (SEQ ID NO: 71), dme-miR-14 (SEQ ID NO: 72), dme-miR-31a (SEQ ID NO: 73), sme-miR-71c (SEQ ID NO: 74), aga-miR-92b (SEQ ID NO: 75), dme-miR-275 (SEQ ID NO: 76), dme-miR-279 (SEQ ID NO: 77), and dme-miR-305 (SEQ ID NO: 78).

3. The method of claim 1 , wherein said recombinant miRNA precursor comprises an RNA sequence selected from the group consisting of SEQ ID NOs:11-18, 36-43, 57-69, 92, and 96.

4. The method of claim 1 , wherein the plant comprises a DNA sequence selected from the group consisting of SEQ ID NOs:1-10, 28-35, 44-46, and 79-88.

5. The method of claim 1 , wherein the invertebrate pest is selected from the group consisting of: a nematode, a Diabrotica species, or a flea beetle.

6. The method of claim 1 , wherein the target gene is a vATPase.

7. The method of claim 1 , wherein the target gene is a cgh-1.

8. A plant comprising a recombinant miRNA precursor, wherein the recombinant miRNA precursor comprises a single strand of RNA that folds into a secondary structure of a miRNA precursor identified from an invertebrate pest genome, wherein said miRNA precursor comprises at least one stem-loop capable of being processed into a mature miRNA capable of suppressing at least one target gene of said invertebrate pest.

9. The plant of claim 8 , wherein the invertebrate pest is selected from the group consisting of: a nematode, a Diabrotica species, or a flea beetle.

10. The plant of claim 8 , wherein the target gene is a vATPase.

11. The plant of claim 8 , wherein the target gene is a cgh-1.

12. The plant of claim 8 , wherein the recombinant miRNA precursor comprises an RNA sequence selected from the group consisting of SEQ ID NOs:11-18, 36-43, 57-69, 92, and 96.

13. The plant of claim 8 , wherein the plant comprises a DNA sequence selected from the group consisting of SEQ ID NOs:1-10, 28-35, 44-46, and 79-88.

14. The plant of claim 8 , wherein the recombinant miRNA precursor comprises a miRNA selected from the group consisting of bta-miR-7 (SEQ ID NO: 70), dme-miR-9a (SEQ ID NO: 71), dme-miR-14 (SEQ ID NO: 72), dme-miR-31a (SEQ ID NO: 73), sme-miR-71c (SEQ ID NO: 74), aga-miR-92b (SEQ ID NO: 75), dme-miR-275(SEQ ID NO: 76), dme-miR-279 (SEQ ID NO: 77), and dme-miR-305(SEQ ID NO: 78).

Assignees

• Monsanto Technology Llc

Inventors

• Allen Edwards
• Donovan William P
• Heck Gregory R
• Roberts James K
• Ursin Virginia
• Zhang Yuanji

Classifications

• C12N15/8285

  for nematode resistance · CPC title

• C12N15/113

  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 title

• C12N2310/14

  interfering nucleic acids [NA] · CPC title

• C12N15/8286 Primary

  for insect resistance · CPC title

• C12N2310/141

  MicroRNAs, miRNAs · CPC title