Improved production of gutted adenovirus in cells expressing adenovirus preterminal protein and DNA polymerase

Dennis Hartigan-O'Connor, Andrea Amalfitano, Jeffrey S. Chamberlain

Research output: Contribution to journalArticlepeer-review

51 Scopus citations

Abstract

Production of gutted, or helper-dependent, adenovirus vectors by current methods is inefficient. Typically, a plasmid form of the gutted genome is transfected with helper viral DNA into 293 cells; the resulting lysate is serially passaged to increase the titer of gutted virions. Inefficient production of gutted virus particles after cotransfection is likely due to suboptimal association of replication factors with the abnormal origins found in these plasmid substrates. To test this hypothesis, we explored whether gutted virus production would be facilitated by transfection into cells expressing various vital replication factors. We observed that C7 cells, coexpressing adenoviral DNA polymerase and preterminal protein, converted plasmid DNA into replicating virus approximately 50 times more efficiently than did 293 cells. This property of C7 cells can be used to greatly increase the efficiency of gutted virus production after cotransfection of gutted and helper vital DNA. These cells should also he useful for generation of recombinant adenovirus from any plasmid-based precursor.

Original languageEnglish (US)
Pages (from-to)7835-7841
Number of pages7
JournalJournal of Virology
Volume73
Issue number9
StatePublished - 1999
Externally publishedYes

ASJC Scopus subject areas

  • Immunology

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