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• W2035594057 abstract "Abstract Background X‐linked severe combined immunodeficiency (SCID‐X1, X‐SCID) is a life‐threatening disease caused by a mutated common cytokine receptor γ chain (γc) gene. Although ex vivo gene therapy, i.e., transduction of the γc gene into autologous CD34 + cells, has been successful for treating SCID‐X1, the retrovirus vector‐mediated transfer allowed dysregulated integration, causing leukemias. Here, to explore an alternative gene transfer methodology that may offer less risk of insertional mutagenesis, we employed the ϕC31 integrase‐based integration system using human T‐cell lines, including the γc‐deficient ED40515(‐). Methods A ϕC31 integrase and a neo r gene expression plasmid containing the ϕC31 attB sequence were co‐delivered by electroporation into Jurkat cells. After G418 selection, integration site analyses were performed using linear amplification mediated‐polymerase chain reaction (LAM‐PCR). ED40515(‐) cells were also transfected with a γc expression plasmid containing attB , and the integration sites were determined. IL‐2 stimulation was used to assess the functionality of the transduced γc in an ED40515(‐)‐derived clone. Results Following co‐introduction of the ϕC31 integrase expression plasmid and the plasmid carrying attB , the efficiency of integration into the unmodified human genome was assessed. Several integration sites were characterized, including new integration sites in intergenic regions on chromosomes 13 and 18 that may be preferred in hematopoietic cells. An ED40515(‐) line bearing the integrated γc gene exhibited stable expression of the γc protein, with normal IL‐2 signaling, as assessed by STAT5 activation. Conclusions This study supports the possible future use of this ϕC31 integrase‐mediated genomic integration strategy as an alternative gene therapy approach for treating SCID‐X1. Copyright © 2006 John Wiley & Sons, Ltd." @default.
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• W2035594057 date "2006-02-28" @default.
• W2035594057 modified "2023-10-17" @default.
• W2035594057 title "Phage φC31 integrase-mediated genomic integration of the common cytokine receptor gamma chain in human T-cell lines" @default.
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• W2035594057 doi "https://doi.org/10.1002/jgm.891" @default.
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