Site-specific, Rep-mediated integration of the intact β-globin locus in the human erythroleukaemic cell line K562

Abstract

The stable, regulated and tissue-specific expression of a therapeutic transgene can be best achieved by the transfer of a complete genomic locus, which will include the short- and long-range regulatory elements that are critical for the accurate control of gene expression. However, when techniques that rely on the random integration of exogenous DNA into the human genome are used for gene transfer, the risk of insertional mutagenesis remains a major issue. Using components derived from the adeno-associated virus (AAV), we have successfully targeted the integration of 200kb bacterial artificial chromosomes containing the entire β-globin locus into the AAVS1 site on human chromosome 19. We sho..