Generation of human artificial vector to allow the reproduction of gene copy number variation

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PhiC31-based recombination allows the multiple introduction of transgenes when the recipient sites(attB sites) are available. We sought to apply this system to the human artificial chromosome(HAC) vector, which can carry the huge size of transgene. We successfully generated the HAC vector carrying multiple copies of attB sites(attB-HAC) in CHO cells. It was confirmed that transgenes tagged with the donor site(attP site) can be loaded onto the attB sites on the attB-HAC, in independent manner of PhiC31 integrase. Intriguingly, Southern blot analyses implicated that multiple copies of transgenes can be loaded onto the attB-HAC in one recombination-reaction in CHO cells. The present study suggests the efficacy of attB-HAC as a tool for reproduction of gene copy number variation in animal models and cell models.