# US 2022/0347288 A1 · Method for producing artificial recombinant rotavirus > Agents: this data is available as JSON from the patentgrep API and MCP server — see https://patentgrep.com/llms.txt - Number: US 2022/0347288 A1 - Document ID: US20220347288A1 - Status: Published application - Kind: A1 - Application number: 17/805,419 - Filed: 2022-06-03 - Published: 2022-11-03 - Earliest claimed: 2016-09-27 - Assignees: Osaka University - Applicants: Osaka University - Inventors: Takeshi Kobayashi; Yuta Kanai - Web page: https://patentgrep.com/patent/US20220347288A1 ## CPC - A61K 39/15: Reoviridae, e.g. calf diarrhea virus - C12N 7/00: Viruses; Bacteriophages; Compositions thereof; Preparation or purification thereof (preparing medicinal viral antigen or antibody compositions, e.g. virus vaccines, A61K39/00) - A61K 39/12: Viral antigens - A61P 31/14: for RNA viruses - C07K 14/005: from viruses - C07K 14/14: Reoviridae, e.g. rotavirus, bluetongue virus, Colorado tick fever virus - C12N 15/09: Recombinant DNA-technology - C12N 15/86: Viral vectors - C12Q 1/701: {Specific hybridization probes} - A61K 2039/5254: {avirulent or attenuated} - C12N 2720/12022: New viral proteins or individual genes, new structural or functional aspects of known viral proteins or genes - C12N 2720/12034: Use of virus or viral component as vaccine, e.g. live-attenuated or inactivated virus, VLP, viral protein - C12N 2720/12051: Methods of production or purification of viral material - C12N 2720/12334: Use of virus or viral component as vaccine, e.g. live-attenuated or inactivated virus, VLP, viral protein - C12N 2720/12351: 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) - A61P: SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS - C07K: PEPTIDES (peptides in foodstuffs A23; obtaining protein compositions for foodstuffs, working-up proteins for foodstuffs A23J; preparations for medicinal purposes A61K; peptides containing beta-lactam rings C07D; cyclic dipeptides not having in their molecule any other peptide link than those which form their ring, e.g. piperazine-2,5-diones, C07D; ergot alkaloids of the cyclic peptide type C07D519/02; macromolecular compounds having statistically distributed amino acid units in their molecules, i.e. when the preparation does not provide for a specific; but for a random sequence of the amino acid units, homopolyamides and block copolyamides derived from amino acids C08G69/00; macromolecular products derived from proteins C08H1/00; preparation of glue or gelatine C09H; single cell proteins, enzymes C12N; genetic engineering processes for obtaining peptides C12N15/00; compositions for measuring or testing processes involving enzymes C12Q; investigation or analysis of biological material G01N33/00) - C12Q: MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS (immunoassay G01N33/53); COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES ## Abstract Provided is a method for producing an artificial recombinant virus of the family Reoviridae, the method comprising the steps of: (1) introducing a FAST protein expression vector and/or a capping enzyme expression vector into host cells; (2) introducing a vector containing expression cassettes for individual RNA genome segments of a virus or introducing a set of single-stranded RNA transcripts from the expression cassettes into host cells; and (3) culturing the host cells. The method of the present invention allows more efficient production of an artificial recombinant virus of the family Reoviridae as compared with conventional methods and allows artificial recombinant rotavirus production without using a helper virus. ## Claims ### Claim 1 (canceled) ### Claim 2 A method for producing an artificial recombinant virus of the family Reoviridae, the method comprising: (1) introducing a FAST protein expression vector into host cells; (2) introducing a vector containing a set of expression cassettes for individual RNA genome segments of a virus or introducing a set of single-stranded RNA transcripts of the RNA genome segments from the expression cassettes into host cells; and (3) culturing the host cells. ### Claim 3 (depends on 2) The method according to claim 2, wherein step (1) further comprises introducing a capping enzyme expression vector into the host cells. ### Claim 4 (depends on 2) The method according to claim 2, wherein the artificial recombinant virus has a mutation introduced in at least one of the RNA genome segments and/or a foreign gene inserted in at least one of the RNA genome segments. ### Claim 5 (depends on 2) The method according to claim 2, wherein the FAST protein is Nelson Bay reovirus p10, Avian reovirus p10, Broome reovirus p13, Reptilian reovirus p14, Baboon reovirus p15, grass carp reovirus p16 or Atlantic salmon reovirus p22. ### Claim 6 (depends on 2) The method according to claim 2, wherein the capping enzyme is a capping enzyme of a DNA or RNA virus which replicates in the cytoplasm of host cells. ### Claim 7 (depends on 2) The method according to claim 2, wherein the expression cassette for an RNA genome segment comprises an RNA polymerase promoter, a DNA encoding the RNA genome segment and a DNA encoding a self-cleaving ribozyme. ### Claim 8 (depends on 7) The method according to claim 7, wherein the RNA polymerase promoter is a T7 promoter, and the host cells are recombinant T7 RNA polymerase-expressing cells. ### Claim 9 (depends on 7) The method according to claim 7, wherein the ribozyme is a hepatitis D virus ribozyme. ### Claim 10 (depends on 2) The method according to claim 2, wherein the host cells are co-cultured with highly virus-susceptible cells. ### Claim 11 (depends on 2) The method according to claim 2, wherein the artificial recombinant virus of the family Reoviridae is an artificial recombinant rotavirus. ### Claim 12 (depends on 11) The method according to claim 11, comprising overexpressing a rotavirus NSP2 gene product and/or a rotavirus NSP5 gene product in the host cells. ### Claim 13 (depends on 11) The method according to claim 11, wherein the artificial recombinant rotavirus expresses a foreign gene, and wherein a vector containing an expression cassette for an RNA genome segment encoding NSP1 which cassette has an insertion of the foreign gene in an NSP1 gene and a 100- to 1550-base deletion in the NSP1 gene is used instead of a vector containing an expression cassette for an RNA genome segment encoding NSP1. ### Claim 14 A method for promoting viral replication, comprising infecting host cells expressing a FAST protein with a virus of the family Reoviridae and culturing the host cells. ### Claim 15 (depends on 14) The method according to claim 14, wherein the virus of the family Reoviridae is a virus of the genus Orthoreovirus or Rotavirus. ### Claim 16 (depends on 14) The method according to claim 14, wherein the FAST protein is selected from Nelson Bay reovirus p10, Avian reovirus p10, Broome reovirus p13, Reptilian reovirus p14, Baboon reovirus p15, grass carp reovirus p16 or Atlantic salmon reovirus p22.