Rescue of DNA-PK Signaling and T-Cell Differentiation by Targeted Genome Editing in a prkdc Deficient iPSC Disease Model

• DOI: 10.1371/journal.pgen.1005239
• Publication Date: 2015-05-22T17:56:02Z
• Authors: Shamim H. Rahman; Johannes Kuehle; Christian Reimann; Tafadzwa Mlambo; Jamal Alzubi; Morgan L. Maeder; Heimo Riedel; Paul Fisch; Tobias Cantz; Cornelia Rudolph; Claudio Mussolino; J. Keith Joung; Axel Schambach; Toni Cathomen
• Keywords: Male; 0301 basic medicine; Genotyping Techniques; T-Lymphocytes; Induced Pluripotent Stem Cells; 610; DNA-Activated Protein Kinase; QH426-470; Mice; 03 medical and health sciences; Genetics; Animals; Humans; Genome; Nuclear Proteins; Cell Differentiation; Zinc Fingers; Fibroblasts; 3. Good health; DNA-Binding Proteins; Disease Models, Animal; HEK293 Cells; Phenotype; NIH 3T3 Cells; Protein Kinases; Research Article; Signal Transduction

In vitro disease modeling based on induced pluripotent stem cells (iPSCs) provides a powerful system to study cellular pathophysiology, especially in combination with targeted genome editing and protocols differentiate iPSCs into affected cell types. this study, we established zinc-finger nuclease-mediated primary fibroblasts generated from mouse model for radiosensitive severe combined immunodeficiency (RS-SCID), rare disorder characterized by sensitivity radiation the absence of lymphocytes due impaired DNA-dependent protein kinase (DNA-PK) activity. Our results demonstrate that gene RS-SCID rescued DNA-PK dependent signaling overcome radiosensitivity. Furthermore, T-cell differentiation was employed stage-specific maturation block causing mutation. Genetic correction restored T-lymphocyte maturation, polyclonal V(D)J recombination receptor followed successful beta-selection. conclusion, provide proof iPSC-based is valuable paradigm SCID modeling, which can be utilized investigate disorders development validate therapy strategies deficiencies. Moreover, emphasizes significance designer nucleases as tool generating isogenic models their future role producing autologous, genetically corrected transplants various clinical applications.