Molecular evidence of genome editing in a mouse model of immunodeficiency
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 cel...
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| Main Authors: | , , , , |
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| Format: | Article (Journal) |
| Language: | English |
| Published: |
29 May 2018
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| In: |
Scientific reports
Year: 2018, Volume: 8 |
| ISSN: | 2045-2322 |
| DOI: | 10.1038/s41598-018-26439-9 |
| Online Access: | Verlag, Volltext: https://doi.org/10.1038/s41598-018-26439-9 Verlag, Volltext: https://www.nature.com/articles/s41598-018-26439-9 
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| Author Notes: | H.H. Abdul-Razak, C.J. Rocca, S.J. Howe, M.E. Alonso-Ferrero, J. Wang, R. Gabriel, C.C. Bartholomae, C.H.V. Gan, M.I. Garín, A. Roberts, M.P. Blundell, V. Prakash, F.J. Molina-Estevez, J. Pantoglou, G. Guenechea, M.C. Holmes, P.D. Gregory, C. Kinnon, C. von Kalle, M. Schmidt, J.A. Bueren, A.J. Thrasher & R.J. Yáñez-Muñoz |
| Summary: | 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. |
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| Item Description: | Gesehen am 02.04.2019 |
| Physical Description: | Online Resource |
| ISSN: | 2045-2322 |
| DOI: | 10.1038/s41598-018-26439-9 |