Integration of functional bacterial artificial chromosomes into human cord lood-derived multipotent stem cells
F Zaibak, J Kozlovski, J Vadolas, JP Sarsero, R Williamson, SE Howden
GENE THERAPY | NATURE PUBLISHING GROUP | Published : 2009
Stem cells from a patient with a genetic disease could be used for cell therapy if it were possible to insert a functional copy of the defective gene. In this study, we investigate the transfection and subsequent integration of large genomic fragments into human cord blood-derived multipotent stem cells. We describe for the first time the creation of clonal stem cells carrying a human bacterial artificial chromosome (BAC) containing the Friedreich ataxia locus with an enhanced green fluorescent protein (EGFP) reporter gene fused to exon 5a of the frataxin (FXN) gene. Integration of the BAC into the host cell genome was confirmed by PCR, Southern blot and fluorescent in situ hybridization ana..View full abstract
We thank the New Zealand Cystic Fibrosis Association, CureKids New Zealand, the Australian Cystic Fibrosis Research Trust and a dedicated team of supporters who initially raised funding that enabled this research to proceed. We are also indebted to the Thalassaemia Society of Victoria, Radiomarathon, Laiki Bank, The Greek Conference and the Greek and Cypriot communities of Melbourne, Australia for their support.