AAV integration in human hepatocytes

Recombinant adeno-associated viral (rAAV) vectors are considered promising tools for gene therapy directed at the liver. While rAAV is thought to be an episomal vector, its single stranded DNA genome is prone to intra and inter -molecular recombination leading to rearrangements and integration into the host cell genome. Here we ascertained the integration frequency of rAAV in human hepatocytes transduced either ex vivo or in vivo and subsequently expanded in a mouse model of xenogeneic liver regeneration. Chromosomal rAAV integration events and vector integrity were determined using the capture-PACBIO sequencing approach, a long-read next- generation sequencing method that has not previously been used for this purpose.