Adeno-associated virus vector gene expression occurs in nondividing cells in the absence of vector DNA integration.

T. R. Flotte, S. A. Afione, P. L. Zeitlin

Research output: Contribution to journalArticle

Abstract

Adeno-associated virus type 2 (AAV2)-based vectors are capable of stable expression in the airway epithelium and may be useful for gene therapy for human diseases, such as cystic fibrosis. Certain virus vectors, such as retroviruses, require active cell division for integration and expression, but this has not been formally evaluated in the case of AAV2. The cystic fibrosis bronchial epithelial cell line, IB3-1, which can be transduced by AAV2 vectors, was shown to undergo a decrease in DNA synthesis to undetectable levels when grown to confluence. Cultures in which <0.1% of cells were dividing could still be efficiently transduced with AAV-lacZ or AAV-neo vectors, with a linear dose response, up to 91% with a multiplicity of 3,000 vector particles per cell. The fate of vector DNA in nondividing target cells was investigated by Southern blotting of both low molecular weight, nonintegrated DNA and high molecular weight, genomic DNA fractions. Detectable levels of vector DNA were only seen in the nonintegrated state. These results indicate that AAV2-based vectors, unlike retrovirus vectors, do not require active cell division or integration for expression to occur and thus possess a unique profile of biologic properties.

Original languageEnglish (US)
Pages (from-to)517-521
Number of pages5
JournalAmerican Journal of Respiratory Cell and Molecular Biology
Volume11
Issue number5
StatePublished - Nov 1994

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Dependovirus
Viruses
Gene expression
Gene Expression
DNA
Retroviridae
Cystic Fibrosis
Cell Division
Molecular Weight
Southern Blotting
Genetic Therapy
Molecular weight
Cells
Epithelium
Epithelial Cells
Gene therapy
Cell Line

ASJC Scopus subject areas

  • Cell Biology
  • Molecular Biology
  • Pulmonary and Respiratory Medicine

Cite this

Adeno-associated virus vector gene expression occurs in nondividing cells in the absence of vector DNA integration. / Flotte, T. R.; Afione, S. A.; Zeitlin, P. L.

In: American Journal of Respiratory Cell and Molecular Biology, Vol. 11, No. 5, 11.1994, p. 517-521.

Research output: Contribution to journalArticle

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