IP Library Granted Patent US 9,901,631
Granted Patent B2
US 9,901,631 · App. 14/371,534 · Granted Feb 27, 2018

Method and cells for the production of viral vaccines

Inventors: Marciela Degrace (Cambridge, MA); Nir Hacohen (Brookline, MA); Sagi Shapira (Boston, MA); Liguo Wu (Quincy, MA)
Assignees: The Broad Institute, Inc.; The General Hospital Corporation
A61K39/145A01K67/0276C12N7/00C12N2760/16034C12N2760/16051C12N2760/16152
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Quick Facts
Patent No.
US 9,901,631
App. No.
14/371,534
Granted
Feb 27, 2018
Kind
B2
Abstract

The present invention provides genetically modified cells useful for viral replication and the production of viral vaccines.

Claims (21)

1. A virus-infected cell selected from the group consisting of Vero cells, baby hamster kidney (BHK) cells, primary chick kidney (PCK) cells, Madin-Darby Canine Kidney (MDCK) cells, Madin-Darby Bovine Kidney cells, 293 cells, COS cells and Human Embryonic Kidney (HEK) 293T cells, which cell produces an increased amount of virus relative to a wild-type cell infected with the same virus and which cell comprises one or more genes selected from the group consisting of HPS5, HPS1, AP3B1, AP3D, SC35, APPBP1, CEBPB, NFE2L2, PDGFRL, PPPIRIC, SFRS2, SNAI2, TAF5L, TJP2, TMEM14C, ZNFF331 and ZNF498 whose expression or activity is reduced or inhibited in said virus-infected cell.

2. The cell of claim 1 , wherein the virus is an influenza virus, an Ebola virus, or a Marburg virus.

3. The cell of claim 1 , wherein the cell is mammalian cell.

4. The cell of claim 3 , wherein the mammalian cell is from a hamster, cattle, monkey, dog or human.

5. A method for replicating a virus comprising: a) providing a cell culture comprising cells selected from the group consisting of Vero cells, baby hamster kidney (BHK) cells, primary chick kidney (PCK) cells, Madin-Darby Canine Kidney (MDCK) cells, Madin-Darby Bovine Kidney cells, 293 cells, COS cells, and Human Embryonic Kidney (HEK) 293T cell, said cells comprising one or more genes selected from the group consisting of HPS5, HPS1, AP3B1, AP3D, SC35, APPBP1, CEBPB, NFE2L2, PDGFRL, PPPIRIC, SFRS2, SNAI2, TAF5L, TJP2, TMEM14C, ZNFF331 and ZNF498, whose expression or activity is reduced or inhibited in said cells;

b) infecting the cells with a virus selected from influenza virus, Ebola virus, Marburg virus, Moloney leukemia virus, Machupo virus, Lassa virus, Lymphocytic choriomeningitis virus, Newcastle disease virus, Vesicular stomatitis virus, or cytomegalovirus virus; and

c) culturing the cells infected in step (b) to replicate the virus; wherein the virus exhibits increased viral replication relative to a wild-type cell.

6. The method of claim 5 , further comprising isolating the virus replicated in step (c).

7. A process of replicating a virus comprising:

a) injecting a fertilized egg with a virus selected from influenza virus, Ebola virus, Marburg virus, Moloney leukemia virus, Machupo virus, Lassa virus, Lymphocytic choriomeningitis virus, Newcastle disease virus, Vesicular stomatitis virus, or cytomegalovirus virus and a compound that inhibits the expression or activity of one or more genes selected from the group consisting of HPS5, HPS1, AP3B1, AP3D, SC35, APPBP1, CEBPB, NFE2L2, PDGFRL, PPPIRIC, SFRS2, SNAI2, TAF5L, TJP2, TMEM14C, ZNFF331 and ZNF498; and

b) incubating the egg for a predetermined period of time to replicate the virus.

8. A process of replicating a virus comprising:

a) injecting a cell with a virus selected from influenza virus, Ebola virus, Marburg virus, Moloney leukemia virus, Machupo virus, Lassa virus, Lymphocytic choriomeningitis virus, Newcastle disease virus, Vesicular stomatitis virus, or cytomegalovirus virus and a compound that inhibits the expression or activity of one or more genes selected from the group consisting of HPS5, HPS1, AP3B1, AP3D, SC35, APPBP1, CEBPB, NFE2L2, PDGFRL, PPPIRIC, SFRS2, SNAI2, TAF5L, TJP2, TMEM14C, ZNFF331 and ZNF498; and

b) incubating the cell for a predetermined period of time to replicate the virus.

9. The process of claim 7 , further comprising isolating the replicated virus in step (b).

10. The process of claim 7 , wherein said compound is a nucleic acid.

11. The process of claim 7 , wherein said compound is a siRNA.

12. The process of claim 8 , wherein the cell comprises a disruption of one or more genes selected from the group consisting of HPS5, HPS1, AP3B1, AP3D, SC35, APPBP1, CEBPB, NFE2L2, PDGFRL, PPPIRIC, SFRS2, SNAI2, TAF5L, TJP2, TMEM14C, ZNFF331 and ZNF498, wherein the disruption results in decreased expression or activity of the one or more genes in the cell.

13. The process of claim 8 , wherein the cell is a vertebrate cell.

14. The process of claim 8 , wherein the cell is a mammalian cell.

15. The process of claim 14 , wherein the mammalian cell is from a hamster, cattle, monkey, dog or human.

Assignments (4)
ASSIGNMENT OF ASSIGNOR'S INTEREST Recorded Apr 13, 2016
From: DEGRACE, MARCIELA
To: THE BROAD INSTITUTE, INC.
Reel/Frame 038270/0192 →
ASSIGNMENT OF ASSIGNOR'S INTEREST Recorded Apr 13, 2016
From: HACOHEN, NIR
To: MASSACHUSETTS GENERAL HOSPITAL
Reel/Frame 038270/0211 →
ASSIGNMENT OF ASSIGNOR'S INTEREST Recorded Mar 13, 2015
From: WU, LIGUO
To: MASSACHUSETTS GENERAL HOSPITAL
Reel/Frame 035159/0172 →
CONFIRMATORY LICENSE Recorded Dec 1, 2014
From: BROAD INSTITUTE, INC.
To: NATIONAL INSTITUTES OF HEALTH (NIH), U.S. DEPT. OF HEALTH AND HUMAN SERVICES (DHHS), U.S. GOVERNMENT
Reel/Frame 034498/0165 →
Continuity (2)
Provisional Application 61585006 · Jan 10, 2012
Related Publication 20150010593A1 · Jan 8, 2015