Production of viruses in avian eggs

Patent metadata

Field	Value
Publication number	US-11118166-B2
Application number	US-201916258099-A
Country	US
Kind code	B2
Filing date	Jan 25, 2019
Priority date	Nov 24, 2015
Publication date	Sep 14, 2021
Grant date	Sep 14, 2021

Abstract

The present invention relates to modified avian eggs which can be used to produce increased levels of virus. The present invention also relates to methods of producing viruses in avian eggs of the invention, as well as the use of the viruses obtained to prepare vaccine compositions.

First claim

The invention claimed is:

1. A method of replicating a virus, the method comprising;
1) inoculating an avian egg with a virus, wherein the avian egg comprises a genetic modification in the genome of the egg which knocks-out expression of the interferon alpha/beta receptor 1 (IFNAR1) gene in the egg, wherein the egg is capable of producing more virus than the isogenic egg lacking the genetic modification; and
2) incubating the egg for a predetermined period of time to replicate the virus.

2. The method of claim 1 , wherein the genetic modification was introduced by a programmable nuclease.

3. The method of claim 2 , wherein the nuclease is selected from a: RNA-guided engineered nuclease (RGEN), transcription activator-like nuclease (TALEN) and zinc-finger nuclease (ZFN).

4. The method of claim 3 , wherein the nuclease is a RNA-guided engineered nuclease (RGEN).

5. The method of claim 2 , wherein the nuclease introduced a deletion, substitution or an insertion into the antiviral gene IFNAR1 gene or a regulatory region thereof.

6. The method of claim 1 , wherein the genetic modification was introduced by homologous recombination.

7. The method of claim 1 , wherein the virus is an animal virus.

8. The method of claim 7 , wherein the virus is in a family selected from:
Orthomyxoviridae, Herpesviridae, Paramyxoviridae, Flaviviridae and Coronaviridae.

9. The method of claim 8 , wherein in the virus in selected from: Influenza virus, Canine distemper virus, Measles virus, Reovirus, Eastern equine encephalitis virus, Canine parainfluenza virus, Rabies virus, Fowlpox virus, Western equine encephalitis virus, Mumps virus, Equine encephalomyelitis, Rubella virus, Egg drop syndrome virus, Avian oncolytic viruses, Avian infectious laryngotracheitis Herpesvirus, Newcastle disease virus, Bovine parainfluenza virus, Smallpox virus, Infectious bursal disease, Bovine Ibaraki virus, Recombinant poxvirus, Avian adenovirus type I, II or III, Swine Japanese encephalitis virus, Yellow fever virus, Herpes virus, Sindbis virus, Infections bronchitis virus, Semliki forest virus, Encephalomyelitis virus, Venezuelan EEV virus, Chicken anaemia virus, Marek's disease virus, Parvovirus, Foot and mouth disease virus, Porcine reproductive and respiratory syndrome virus, Classical swine fever virus, Bluetongue virus, Kabane virus, Infectious salmon anaemia virus, Infectious hematopoietic necrosis virus, Viral haemorrhagic septicemia virus and Infectious pancreatic necrosis virus.

10. The method of claim 9 , wherein the virus is an Influenza virus.

11. The method of claim 1 , wherein the egg is a chicken egg.

12. The method of claim 1 which further comprises harvesting the replicated virus.

13. The method of claim 12 , wherein the replicated virus are harvested from allantoic fluid of the egg.

14. The method of claim 1 , wherein following step 2) the replicated virus are inactivated.

15. The method of claim 1 , wherein following step 2) the replicated virus are formulated into a vaccine composition.

16. The method of claim 7 , wherein the virus is in a family Orthomyxoviridae.

17. The method of claim 7 , wherein the animal virus is a human virus.

18. The method of claim 1 , wherein the egg is capable of producing at least 1.5 fold more virus than the isogenic egg.

19. The method of claim 1 , wherein the egg is capable of producing at least 2 fold more virus than the isogenic egg.

Assignees

• Commw Scient Ind Res Org
• Univ Georgia

Inventors

• Bean Andrew
• Lowenthal John William
• Malaver-Ortega Luis Fernando
• Tripp Ralph A

Classifications

• C12N7/00 Primary

  Viruses; Bacteriophages; Compositions thereof; Preparation or purification thereof (preparing medicinal viral antigen or antibody compositions, e.g. virus vaccines, A61K39/00) · CPC title

• A01K2227/30

  Bird · CPC title

• A01K67/0275

  Genetically modified vertebrates, e.g. transgenic · CPC title

• C12N2760/16151

  Methods of production or purification of viral material · CPC title

• C12N15/1133

  against herpetoviridae, e.g. HSV · CPC title