Molecular Evidence of Genome Editing in a Mouse Model of Immunodeficiency. / Abdul-Razak, H. H.; Rocca, C. J.; Howe, S. J.; Alonso-Ferrero, M. E.; Wang, J.; Gabriel, R.; Bartholomae, C. C.; Gan, C. H.V.; Garín, M. I.; Roberts, A.; Blundell, M. P.; Prakash, V.; Molina-Estevez, F. J.; Pantoglou, J.; Guenechea, G.; Holmes, M. C.; Gregory, P. D.; Kinnon, C.; Von Kalle, C.; Schmidt, M.; Bueren, J. A.; Thrasher, A. J.; Yáñez-Muñoz, R. J.
In: Scientific Reports, Vol. 8, 8214, 29.05.2018, p. 1-13.Research output: Contribution to journal › Article › peer-review
Genome editing is the introduction of directed modifications in the genome, a process boosted to therapeutic levels by designer nucleases. Building on the experience of ex vivo gene therapy for severe combined immunodeficiencies, it is likely that genome editing of haematopoietic stem/progenitor cells (HSPC) for correction of inherited blood diseases will be an early clinical application. We show molecular evidence of gene correction in a mouse model of primary immunodeficiency. In vitro experiments in DNA-dependent protein kinase catalytic subunit severe combined immunodeficiency (Prkdc scid) fibroblasts using designed zinc finger nucleases (ZFN) and a repair template demonstrated molecular and functional correction of the defect. Following transplantation of ex vivo gene-edited Prkdc scid HSPC, some of the recipient animals carried the expected genomic signature of ZFN-driven gene correction. In some primary and secondary transplant recipients we detected double-positive CD4/CD8 T-cells in thymus and single-positive T-cells in blood, but no other evidence of immune reconstitution. However, the leakiness of this model is a confounding factor for the interpretation of the possible T-cell reconstitution. Our results provide support for the feasibility of rescuing inherited blood disease by ex vivo genome editing followed by transplantation, and highlight some of the challenges.
Original language | English |
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Article number | 8214 |
Pages (from-to) | 1-13 |
Number of pages | 13 |
Journal | Scientific Reports |
Volume | 8 |
DOIs | |
Publication status | Published - 29 May 2018 |
ID: 30428242