Nature Medicine8, 1166 - 1170 (2002)
Published online: 16 September 2002; Corrected online: 23 September 2002 | doi:10.1038/nm766
There is a Corrigendum (February 2003) associated with this Article.
Stable nonviral genetic correction of inherited human skin disease
Susana Ortiz-Urda1, Bhaskar Thyagarajan2, Douglas R. Keene3, Qun Lin1, Min Fang1, Michele P. Calos2
& Paul A. Khavari1
1
VA Palo Alto Healthcare System, Palo Alto, California and the Program in Epithelial Biology, Stanford University School of Medicine, Stanford, California, USA
2
Department of Genetics, Stanford University School of Medicine, Stanford, California, USA
3
Shriners Hospital for Children, Portland, Oregon, USA
Current gene-transfer technologies display limitations in achieving effective gene delivery. Among these limitations are difficulties in stably integrating large corrective sequences into the genomes of long-lived progenitor-cell populations. Current larger-capacity viral vectors suffer from biosafety concerns, whereas plasmid-based approaches have poor efficiency of stable gene transfer1,
2. These barriers hinder genetic correction of many severe inherited human diseases, such as the blistering skin disorder recessive dystrophic epidermolysis bullosa (RDEB)3, caused by mutations in the large COL7A1 gene. To circumvent these barriers, we used the C31 bacteriophage integrase4,
5,
6, which stably integrates large DNA sequences containing a specific 285-base-pair attB sequence into genomic 'pseudo-attP sites'. C31 integrase−based gene transfer stably integrated the COL7A1 cDNA into genomes of primary epidermal progenitor cells from four unrelated RDEB patients. Skin regenerated using these cells displayed stable correction of hallmark RDEB disease features, including Type VII collagen protein expression, anchoring fibril formation and dermal-epidermal cohesion. These findings establish a practical approach to nonviral genetic correction of severe human genetic disorders requiring stable genomic integration of large DNA sequences.
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C31 bacteriophage integrase
B in growth factor regulated type VII collagen gene expression
