Characterization of a preferred site on human chromosome 19q for integration of adeno‐associated virus DNA by non‐homologous recombination

• Published in: The EMBO journal Vol. 11; no. 13; pp. 5071 - 5078
• Main Authors: Kotin, R.M., Linden, R.M., Berns, K.I.
• Format: Journal Article
• Language: English
• Published: England 01-12-1992
• Subjects: adeno-associated virus; Base Sequence; chromosome 19; Chromosomes, Human, Pair 19; Cloning, Molecular; Dependovirus - physiology; DNA; DNA, Viral - metabolism; Humans; integration; man; Molecular Sequence Data; Nucleic Acid Conformation; Polymerase Chain Reaction; Promoter Regions, Genetic; recombination; Recombination, Genetic; TATA Box; Transcription, Genetic; Virus Integration
• ISSN: 0261-4189; 1460-2075
• DOI: 10.1002/j.1460-2075.1992.tb05614.x

The human parvovirus, adeno‐associated virus (AAV), has been shown to integrate preferentially into human chromosome 19 q13.3‐qter. The human target sequence for AAV integration (AAVS1) was cloned and sequenced. By analysis of the proviral junctions it was determined that integration of the AAV DNA occurred via a non‐homologous recombination pathway although there were either four or five identical nucleotides at the junctions. Integration was a multistep, concerted process that resulted in cellular sequence rearrangements. The sequence of the integration locus was analyzed for possible recombination signals. Direct repeats at a much greater than random occurrence were found distributed non‐uniformly throughout the AAVS1 sequence. A CpG island containing transcription factor binding site elements is suggestive of a TATA‐less promoter. Evidence for transcriptional activity was provided by PCR amplification of reverse transcribed RNA.