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Related Experiment Videos

Site-specific integration by adeno-associated virus.

R M Kotin1, M Siniscalco, R J Samulski

  • 1Hearst Microbiology Research Center, Department of Microbiology, Cornell University Medical College, New York, NY 10021.

Proceedings of the National Academy of Sciences of the United States of America
|March 1, 1990
PubMed
Summary
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The adeno-associated virus (AAV) genome shows a preference for integrating into a specific region on human chromosome 19. This targeted integration was identified across multiple independently derived infected cell clones.

Area of Science:

  • Molecular Biology
  • Virology
  • Genetics

Background:

  • Adeno-associated virus (AAV) is a small DNA virus with potential therapeutic applications.
  • Understanding AAV integration into the host genome is crucial for assessing its safety and efficacy.
  • Latent infections involve the stable presence of viral genomes within host cells.

Purpose of the Study:

  • To investigate the integration patterns of the adeno-associated virus (AAV) genome in human cells.
  • To identify specific cellular genomic locations targeted by AAV integration.
  • To characterize the chromosomal localization of AAV integration sites.

Main Methods:

  • Isolation of cellular sequences flanking integrated AAV genomes from infected cell lines.
  • Genomic blotting using flanking-sequence and AAV probes on DNA from multiple infected clones.

Related Experiment Videos

  • Somatic cell hybrid mapping to determine chromosomal localization of integration sites.

    • Main Results:

    • AAV integration was observed in 15 out of 22 independently derived latently infected human cell clones.
    • Hybridization patterns indicated preferential integration into specific cellular genomic regions.
    • Somatic cell hybrid mapping identified chromosome 19 as the unique integration site for AAV.

    Conclusions:

    • The adeno-associated virus (AAV) genome demonstrates a strong preference for integrating into a specific locus within the human genome.
    • This preferential integration site has been mapped to chromosome 19.
    • The identified AAV integration site sequence has been successfully isolated from uninfected human DNA.