Site-specific, Rep-mediated integration of the intact beta-globin locus in the human erythroleukaemic cell line K562
SE Howden, L Voullaire, H Wardan, R Williamson, J Vadolas
GENE THERAPY | NATURE PUBLISHING GROUP | Published : 2008
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 200 kb bacterial artificial chromosomes containing the entire beta-globin locus into the AAVS1 site on human chromosome 19. We..View full abstract
We thank Dr Horace Drew and Dr Gerry for providing the p5-22 and pRep10 plasmids, and for their helpful advice. We also thank Dr Jude Samulski for providing the pHIV68 plasmid. This work was supported by the National Health and Medical Research Council, the Murdoch Children's Research Institute and the Thalassaemia Society of Victoria.