# US 10,369,210 B2 · Cells for producing influenza virus and method for producing influenza virus > Agents: this data is available as JSON from the patentgrep API and MCP server — see https://patentgrep.com/llms.txt - Number: US 10,369,210 B2 - Document ID: US10369210B2 - Status: Granted - Kind: B2 - Application number: 15/511,988 - Filed: 2015-09-18 - Published: 2018-04-19 - Granted: 2019-08-06 - Earliest claimed: 2014-09-22 - Assignees: Japan Science and Technology Agency - Applicants: Japan Science and Technology Agency - Inventors: Yoshihiro Kawaoka; Tokiko Watanabe; Eiryo Kawakami; Shinji Watanabe - Web page: https://patentgrep.com/patent/US10369210B2 ## CPC - A61K 39/145: Orthomyxoviridae, e.g. influenza virus - C12N 15/111: {General methods applicable to biologically active non-coding nucleic acids} - C12N 5/00: Undifferentiated human, animal or plant cells, e.g. cell lines; Tissues; Cultivation or maintenance thereof; Culture media therefor; (plant reproduction by tissue culture techniques A01H4/00) - C12N 5/0686: {Kidney cells} - C12N 5/10: Cells modified by introduction of foreign genetic material - C12N 7/00: Viruses; Bacteriophages; Compositions thereof; Preparation or purification thereof (preparing medicinal viral antigen or antibody compositions, e.g. virus vaccines, A61K39/00) - C12N 7/02: Recovery or purification - C12N 7/08: {Inactivation or attenuation} by serial passage of virus - 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 2310/14: interfering nucleic acids [NA] - C12N 2320/12: in functional genomics, i.e. for the determination of gene function - C12N 2510/00: Genetically modified cells - C12N 2510/02: Cells for production - C12N 2760/16051: Methods of production or purification of viral material - C12N 2760/16151: Methods of production or purification of viral material - 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) - C12N: MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA (microbiological testing media C12Q1/00) ## Abstract The present invention provides cells which have a high ability to propagate influenza virus, are suitable for use in production of an influenza virus for preparing a vaccine, and are able to be cultured in vitro, and a method for producing an influenza virus using the cells. That is, the present invention provides cells for producing an influenza virus in which expression of one or more genes that encode proteins involved in an effect of suppressing influenza virus production in a cell is suppressed and the gene is at least one selected from the group including ACTG1 gene and the like, and a method for producing an influenza virus that includes infecting the cells for producing an influenza virus with an influenza virus and then culturing. ## Claims ### Claim 1 A method for producing an influenza virus, the method comprising: infecting cells with an influenza virus, thereby producing infected cells and culturing the infected cells, wherein the cells are HEK293 cells, Vero cells, or MDCK cells in which expression of at least one chromosomal gene is suppressed and the cells are able to be cultured in vitro, and wherein the at least one chromosomal gene is at least one selected from the group consisting of ACTG1 gene, ACTN1 gene, CCT6A gene, COPS7B gene, DAP3 gene, ERLIN2 gene, GNAI2 gene, GTF3C5 gene, HNRPAB gene, KHSRP gene, KRT18 gene, MTCH1 gene, PPP2R1A gene, PRPS1 gene, RAB18 gene, RAB3A gene, RAP1B gene, RPS19 gene, RPS7 gene, S100A10 gene, SFRS7 gene, SLC2A12 gene, TMED2 gene and USP10 gene. ### Claim 2 (depends on 1) The method for producing an influenza virus according to claim 1, wherein the influenza virus is formulated into an influenza virus vaccine. ### Claim 3 (depends on 1) The method for producing an influenza virus according to claim 1, wherein the at least one chromosomal gene is deleted or disrupted. ### Claim 4 (depends on 1) The method for producing an influenza virus according to claim 1, wherein the at least one chromosomal gene is at least one selected from the group including ACTG1 gene, ACTN1 gene, DAP3 gene, GNAI2 gene, GTF3C5 gene, KRT18 gene, MTCH1 gene, PRPS1 gene, RPS19 gene, S100A10 gene, SFRS7 gene, SLC2A12 gene, USP10 gene, PPP2R1A gene, and TMED2 gene. ### Claim 5 (depends on 1) The method for producing an influenza virus according to claim 1, wherein the cells are HEK 293 cells. ### Claim 6 (depends on 5) The method for producing an influenza virus according to claim 5, the method further comprising: introducing an siRNA for the at least one chromosomal gene to the cells prior to the infecting. ### Claim 7 (depends on 1) The method for producing an influenza virus according to claim 1, wherein the cells are Vero cells. ### Claim 8 (depends on 7) The method for producing an influenza virus according to claim 7, wherein the at least one chromosomal gene is at least one selected from the group consisting of ACTG1 gene, ACTN1 gene, DAP3 gene, GNAI2 gene, GTF3C5 gene, KRT18 gene, MTCH1 gene, PRPS1 gene, RPS19 gene, S100A10 gene, SFRS7 gene, SLC2A12 gene and USP10 gene. ### Claim 9 (depends on 8) The method for producing an influenza virus according to claim 8, the method further comprising: introducing an siRNA for the at least one chromosomal gene to the cells prior to the infecting. ### Claim 10 (depends on 1) The method for producing an influenza vines according to claim 1, wherein the cells are MDCK cells. ### Claim 11 (depends on 10) The method for producing an influenza virus according to claim 10, wherein the at least one chromosomal gene is at least one selected from the group consisting of KRT18 gene, PPP2R1A gene, TMED2 gene and USP10 gene. ### Claim 12 (depends on 11) The method for producing an influenza virus according to claim 11, the method further comprising: introducing an siRNA for the at least one chromosomal gene to the cells prior to the infecting. ### Claim 13 (depends on 10) The method for producing an influenza virus according to claim 10, wherein the at least one chromosomal gene is at least one selected from the group consisting of KRT18 gene and USP10 gene. ### Claim 14 (depends on 11) The method for producing an influenza virus according to claim 11, the method further comprising: introducing CRISPR/Cas system comprising at least one guide RNA that recognizes a target sequence to be cut in the at least one chromosomal gene to the cells prior to the infecting.