# US 8,772,254 B2 · Method and constructs for delivering double stranded RNA to pest organisms

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- Number: US 8,772,254 B2
- Document ID: US8772254B2
- Status: Granted
- Kind: B2
- Application number: 11/666,017
- Filed: 2005-10-25
- Published: 2009-10-22
- Granted: 2014-07-08
- Earliest claimed: 2004-10-25
- Assignees: Devgen NV
- Applicants: Thierry AndréOlivier Eddy Bogaert; Richard Zwaal; Geert Karel Maria Plaetinck; Jan Octaaf De Kerpel; Titus Jan Kaletta
- Inventors: Thierry AndréOlivier Eddy Bogaert; Richard Zwaal; Geert Karel Maria Plaetinck; Jan Octaaf De Kerpel; Titus Jan Kaletta
- Web page: https://patentgrep.com/patent/US8772254B2

## CPC

- C12N 15/8279: {for biotic stress resistance, pathogen resistance, disease resistance}
- A61K 48/0033: {the non-active part being non-polymeric}
- C12N 15/111: {General methods applicable to biologically active non-coding nucleic acids}
- 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)}
- C12N 15/8216: {Methods for controlling, regulating or enhancing expression of transgenes in plant cells}
- C12N 15/8283: {for virus resistance}
- C12N 15/8285: {for nematode resistance}
- C12N 15/8286: {for insect resistance}
- A01K 2217/058: due to expression of inhibitory nucleic acid, e.g. siRNA, antisense
- A01K 2227/703: Worms, e.g. Caenorhabdities elegans
- C12N 2310/14: interfering nucleic acids [NA]
- C12N 2310/3513: Protein; Peptide
- C12N 2320/32: Special delivery means, e.g. tissue-specific
- C12N 2710/14143: viral genome or elements thereof as genetic vector
- C12N 2770/00011: Details
- Y02A 40/146: Genetically Modified [GMO] plants, e.g. transgenic plants
- C12N: MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA (microbiological testing media C12Q1/00)
- A61K: PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES (devices or methods specially adapted for bringing pharmaceutical products into particular physical or administering forms A61J3/00; chemical aspects of, or use of materials for deodorisation of air, for disinfection or sterilisation, or for bandages, dressings, absorbent pads or surgical articles A61L; soap compositions C11D)
- A01K: ANIMAL HUSBANDRY; AVICULTURE; APICULTURE; PISCICULTURE; FISHING; REARING OR BREEDING ANIMALS, NOT OTHERWISE PROVIDED FOR; NEW BREEDS OF ANIMALS
- Y02A: TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE

## Abstract

The present invention relates in generally to RNAi and its use in gene silencing. In particular, the present invention relates to methods and constructs for delivering double stranded RNA (dsRNA) to pest organisms.

## Claims

### Claim 1

A method for delivering a double-stranded RNA molecule to a pest species, comprising
co-expressing in a plant cell of:
a) an RNA delivery molecule consisting of a polypeptide sequence comprising
i) at least one sequence-specific RNA-binding domain,
ii) at least one targeting polypeptide, able to bind to a cellular endocytosis and/or transcytosis receptor molecule, and
iii) optionally at least one peptide linker for linking the sequence-specific RNA binding domain (i) to the targeting polypeptide (ii), and
b) an RNA molecule which comprises double-stranded RNA comprising annealed complementary strands, one of which has a nucleotide sequence which is complementary to at least part of a target nucleotide sequence of a target gene of a pest species, and which further comprises a nucleotide sequence which specifically binds to the sequence-specific RNA-binding domain of the RNA delivery molecule of (a); and
feeding said plant cell to said pest species.

### Claim 2 (depends on 1)

The method of claim 1, wherein the nucleotide sequence which binds to the sequence-specific RNA-binding domain of the RNA delivery molecule of (a) is a nucleotide sequence as provided by SEQ ID NO:40.

### Claim 3 (depends on 1)

The method of claim 1, wherein the sequence-specific RNA-binding domain of (a)(i) is a polypeptide comprising the bacteriophage MS2 coat protein as represented in SEQ ID NO:14.

### Claim 4 (depends on 1)

The method of claim 1, wherein the targeting polypeptide of (a)(ii) is a snowdrop lectin.