# US 2014/0193858 A1 · Production of viral vectors > Agents: this data is available as JSON from the patentgrep API and MCP server — see https://patentgrep.com/llms.txt - Number: US 2014/0193858 A1 - Document ID: US20140193858A1 - Status: Published application - Kind: A1 - Application number: 14/163,331 - Filed: 2014-01-24 - Published: 2014-07-10 - Earliest claimed: 2000-09-25 - Assignees: The Regents of the University of Michigan - Applicants: The Regents of the University of Michigan - Inventors: Jeffrey S. CHAMBERLAIN; Dennis J. HARTIGAN-O'CONNOR - Web page: https://patentgrep.com/patent/US20140193858A1 ## CPC - C12N 15/86: Viral vectors - C12N 2710/10043: viral genome or elements thereof as genetic vector - 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 2710/10351: Methods of production or purification of viral material - C12N 2800/30: Vector systems comprising sequences for excision in presence of a recombinase, e.g. loxP or FRT - C12N 2830/002: inducible enhancer/promoter combination, e.g. hypoxia, iron, transcription factor - C12N 2830/38: being a stuffer - 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 relates to methods and compositions for the production of viral vectors. In particular, the present invention provides methods and compositions for faster, higher titer and higher purity production of viral vectors (e.g. adenoviral vectors). In some embodiments, the present invention provides gutted and helper viruses with identical or similar termini. In other embodiments, the present invention provides terminal protein linked adenoviral DNA. In certain embodiments, the present invention provides template extended adenoviral DNA. ## Claims ### Claim 1 A method for producing helper-dependent viral vectors comprising: a) providing; i) helper-dependent viral DNA comprising a first origin of replication, ii) helper viral DNA comprising a second origin of replication, wherein said second origin of replication has similar activity level in a replication assay as said first origin of replication, and iii) target cells; and b) transfecting said target cells with said helper-dependent viral DNA and said helper viral DNA under conditions such that helper-dependent viral vectors are produced. ### Claim 2 (depends on 1) The method of claim 1, wherein said helper-dependent viral DNA comprises adenoviral DNA. ### Claim 3 (depends on 1) The method of claim 1, wherein said helper-dependent viral DNA comprises a heterologous gene sequence. ### Claim 4 (depends on 1) The method of claim 1, wherein said first origin of replication and said second origin of replication have nucleic acid sequences that are substantially similar. ### Claim 5 (depends on 1) The method of claim 1, wherein said helper viral DNA is adenoviral helper viral DNA. ### Claim 6 (depends on 1) The method of claim 1, wherein said first origin of replication and said second origin of replication are not linked to terminal protein or any terminal protein remnant. ### Claim 7 (depends on 1) The method of claim 1, wherein said helper viral DNA comprises a crippling sequence. ### Claim 8 (depends on 1) The method of claim 1, wherein said helper viral DNA comprises recognition sites for site-specific recombinases. ### Claim 9 (depends on 1) The method of claim 1, wherein said target cells express adenoviral DNA polymerase and preterminal protein. ### Claim 10 (depends on 8) The method of claim 8, further comprising; providing iv) a vector encoding a site-specific recombinase, and step c) transfecting said target cells with said vector. ### Claim 11 (depends on 10) The method of claim 10, further comprising recovering said helper-dependent vectors. ### Claim 12 (depends on 10) The method of claim 10, wherein said recovering yields a helper-dependent titer of at least 20 fold increase compared to transfection/infection protocols in cells expressing adenoviral DNA polymerase and preterminal protein. ### Claim 13 (depends on 1) A composition comprising said helper-dependent viral vectors produced by the method of claim 1. ### Claim 14 A host cell comprising; a) helper-dependent viral DNA comprising a first origin of replication, and b) helper viral DNA comprising a second origin of replication, wherein said second origin of replication has a similar activity level in a replication assay as said first origin of replication. ### Claim 15 A method for producing helper-dependent viral vectors comprising: a) providing; i) helper-dependent viral DNA comprising an origin of replication linked to a replication-promoting agent, and ii) target cells; and b) transfecting said target cells with said helper-dependent viral DNA under conditions such that helper-dependent viral vectors are produced. ### Claim 16 (depends on 15) The method of claim 15, further comprising; providing iii) helper viral DNA, and step c) transfecting said target cells with said helper viral DNA. ### Claim 17 (depends on 15) The method of claim 15, wherein said replication-promoting agent is selected from Ad2 preterminal protein, Ad2 terminal protein, Ad5 preterminal protein, and Ad5 terminal protein. ### Claim 18 (depends on 15) The method of claim 15, wherein said helper-dependent viral DNA comprises adenoviral DNA. ### Claim 19 (depends on 15) The method of claim 15, wherein said helper-dependent viral DNA comprises a heterologous gene sequence. ### Claim 20 (depends on 15) The method of claim 15, wherein said helper viral DNA is linked to adenoviral terminal protein.