# US 6,127,175 A · Cells for the production of recombinant adenoviruses > Agents: this data is available as JSON from the patentgrep API and MCP server — see https://patentgrep.com/llms.txt - Number: US 6,127,175 A - Document ID: US6127175A - Status: Granted - Kind: A - Application number: 08/875,223 - Filed: 1997-07-17 - Granted: 2000-10-03 - Earliest claimed: 1995-01-20 - Assignees: Rhone-Poulenc Rorer S.A. - Inventors: Emmanuelle Vigne; Michel Perricaudet; Jean-Fran.cedilla.ois Dedieu; Cecile Orsini; Patrice Yeh; Martine Latta; Edouard Prost - Web page: https://patentgrep.com/patent/US6127175A ## CPC - C12N 15/86: Viral vectors - 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 2710/10322: New viral proteins or individual genes, new structural or functional aspects of known viral proteins or genes - C12N 2710/10343: viral genome or elements thereof as genetic vector - C12N 2710/10362: by genetic engineering - C12N 2750/14143: viral genome or elements thereof as genetic vector - C12N 2830/002: inducible enhancer/promoter combination, e.g. hypoxia, iron, transcription factor - C12N: MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA (microbiological testing media C12Q1/00) ## Abstract The invention relates to cells usable for the production of defective adenoviruses comprising, inserted into their genome, a portion of the region E4 of an adenovirus genome carrying the reading phase ORF6 under the control of a functional promoter. ## Claims ### Claim 1 A recombinant cell line for the production of a defective adenovirus comprising, inserted into its genome, part of an adenovirus E4 region comprising an ORF6 reading frame under the control of a functional promoter, wherein the inserted E4 region does not contain a functional ORF4 reading frame. ### Claim 2 (depends on 1) The cell line according to claim 1, wherein the E4 region further comprises a reading frame ORF6/7. ### Claim 3 (depends on 1) The cell line according to claim 1, wherein the E4 region is derived from a group C human adenovirus genome. ### Claim 4 (depends on 3) The cell line according to claim 3, wherein the E4 region is derived from the genome of an Ad2 or Ad5 adenovirus. ### Claim 5 (depends on 1) The cell line according to claim 1, wherein the promoter is an inducible promoter. ### Claim 6 (depends on 5) The cell line according to claim 5, wherein the promoter is an MMTV promoter. ### Claim 7 (depends on 1) The cell line according to claim 1, wherein the reading frame ORF6 is fused in translational phase with a domain of a nuclear receptor responsible for recognizing its specific ligand. ### Claim 8 (depends on 7) The cell line according to claim 7, wherein the nuclear receptor domain is a hormone-binding domain of the glucocorticoid receptor (HBD-GCR). ### Claim 9 (depends on 8) The cell line according to claim 8, wherein the reading frame ORF6 fused in translational phase with the domain of a nuclear receptor responsible for recognizing its specific ligand is placed under the control of an inducible promoter which responds to a glucocorticoid. ### Claim 10 (depends on 9) The cell line according to claim 9, wherein the inducible promoter is a GRE5 promoter. ### Claim 11 (depends on 1) The cell line according to claim 1, which transcomplements for the E1 region. ### Claim 12 (depends on 11) The cell line according to claim 11, which is derived from cell line 293. ### Claim 13 (depends on 1) The cell line according to claim 1, which is derived from a cell line selected from the group consisting of KB, Hela, MDCK, Vero, or gmDBP6 cells. ### Claim 14 (depends on 1) The cell line according to claim 1, which is derived from a culture of primary cells. ### Claim 15 (depends on 1) The cell line according to claim 1, wherein the part of the E4 region does not contain ORF4. ### Claim 16 (depends on 15) The cell line according to claim 15, wherein the part of the E4 region does not contain ORF1-ORF4. ### Claim 17 (depends on 1) The cell line according to claim 1, wherein the part of the E4 region is a BglII-BglII fragment corresponding to nucleotides 34115-32490 of the Ad5 genome, or the corresponding nucleotides from Ad2, Ad7 or Ad 12. ### Claim 18 (depends on 17) The cell line according to claim 17, which is a cell of a 293 line. ### Claim 19 (depends on 1) The cell line according to claim 1, wherein the part of the E4 region is a BglII-PvuII fragment corresponding to nucleotides 34115-33126 of the Ad5 genome, or the corresponding nucleotides from Ad2, Ad7 or Ad12. ### Claim 20 A plasmid comprising part of an E4 region of an adenovirus genome carrying a reading frame ORF6 under the control of an inducible promoter. ### Claim 21 (depends on 20) The plasmid according to claim 20, wherein the E4 region further comprises a reading frame ORF6/7. ### Claim 22 (depends on 20) The plasmid according to claim 20, wherein the reading frame ORF6 is fused in translational phase with a domain of a nuclear receptor responsible for recognizing its specific ligand. ### Claim 23 (depends on 1) A method for the production of a recombinant adenovirus which is defective at least for the E4 region, comprising infecting the cell line of claim 1 with the E4 defective adenovirus and harvesting the virus. ### Claim 24 (depends on 23) The method according to claim 23, wherein the cell line cells are transformed with one or more plasmids providing the various regions of the genome of the defective recombinant adenovirus. ### Claim 25 (depends on 24) The method according to claim 24, wherein the recombinant adenovirus is defective for E1 and E4 regions. ### Claim 26 A defective recombinant adenovirus .DELTA.E1,ORF3.sup.+,ORF6.sup.-, wherein all or part of the E1 region and of nucleotides 34801-34329 and 34115-33126 of the Ad5 E4 region, or the corresponding nucleotides from Ad2, Ad7 or Ad12, are deleted. ### Claim 27 A defective recombinant adenovirus .DELTA.E1,.DELTA.E4,ORF1.sup.+, wherein all or part of the E1 region and the E4 region with the exception of reading frame ORF1 are deleted. ### Claim 28 (depends on 27) The defective recombinant adenovirus according to claim 27, wherein a fragment whose 5' end is contained in the reading frame ORF7 and whose 3' end is situated in the reading frame ORF2 is deleted. ### Claim 29 (depends on 28) The defective recombinant adenovirus according to claim 28, wherein the Ad5 E4 region covering nucleotides 33093 to 35053, or the corresponding region from Ad2, Ad7 or Ad12, is deleted. ### Claim 30 A defective recombinant adenovirus .DELTA.E1,.DELTA.E4,ORF4.sup.+, wherein all or part of the E1 region and the E4 region with the exception of the reading frame ORF4 are deleted. ### Claim 31 (depends on 30) The defective recombinant adenovirus according to claim 30, wherein the E1 region, a fragment whose 5' end is contained in the reading frame ORF7 and whose 3' end is situated in the reading frame ORF6, and a fragment whose 5' end is contained in the reading frame ORF3 and whose 3' end is situated in the reading frame ORF1 or in the E4 promoter region are deleted. ### Claim 32 (depends on 31) The defective recombinant adenovirus according to claim 31, wherein nucleotides 33093(Smal)-33695 and nucleotides 34634-35355(SmaI) of the Ad5 E4 region, or the corresponding nucleotides from Ad2, Ad7 or Ad12, are deleted. ### Claim 33 A defective recombinant adenovirus .DELTA.E1,.DELTA.E4, wherein all or part of the E1 region and the whole of the E4 region, chosen from the group consisting of Ad5 nucleotides 33466-35355 and 33093-35355, or the corresponding nucleotides from Ad2, Ad7 or Ad12, are deleted.