Delayed expression of adeno-associated virus vector DNA

Sandra A. Afione, Jianming Wang, Scott Walsh, William B. Guggino, Terence R. Flotte

Research output: Contribution to journalArticlepeer-review

Abstract

Two previous reports indicated that recombinant adeno-associated virus (rAAV) vectors were dependent on helper adenovirus (Ad) for efficient conversion of single-stranded (ss) rAAV DNA to the double-stranded (ds) form. This finding is somewhat paradoxical, however, since during a latent infection wild-type (wt)-AAV is rapidly converted to a ds form in the absence of Ad. Our hypothesis was that the effect observed in the previous studies was due to kinetic factors, i.e. to a relative delay in conversion to ds-DNA rather than to an absolute requirement for Ad. To test this, Hela cells were infected with a rAAV-CMV-green fluorescent protein (GFP) vector either in the presence or absence of Ad. Within the first 2 days, Ad infection resulted in a 4-fold increase in AAV vector expression and an augmentation of conversion to a ds-AAV DNA. By 6 days, however, the total number of GFP-expressing cells in the Ad-free culture had exceeded the original number in the Ad co-infected cells, and the conversion to ds-DNA episomes was substantial and ongoing.

Original languageEnglish (US)
Pages (from-to)213-220
Number of pages8
JournalIntervirology
Volume42
Issue number4
DOIs
StatePublished - Nov 1999

Keywords

  • Adeno-associated virus
  • Adenovirus
  • Gene therapy

ASJC Scopus subject areas

  • Virology
  • Infectious Diseases

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