Targeted genome editing restores T cell differentiation in a humanized X-SCID pluripotent stem cell disease model

Sci Rep. 2017 Sep 29;7(1):12475. doi: 10.1038/s41598-017-12750-4.


The generation of T cells from pluripotent stem cells (PSCs) is attractive for investigating T cell development and validating genome editing strategies in vitro. X-linked severe combined immunodeficiency (X-SCID) is an immune disorder caused by mutations in the IL2RG gene and characterised by the absence of T and NK cells in patients. IL2RG encodes the common gamma chain, which is part of several interleukin receptors, including IL-2 and IL-7 receptors. To model X-SCID in vitro, we generated a mouse embryonic stem cell (ESC) line in which a disease-causing human IL2RG gene variant replaces the endogenous Il2rg locus. We developed a stage-specific T cell differentiation protocol to validate genetic correction of the common G691A mutation with transcription activator-like effector nucleases. While all ESC clones could be differentiated to hematopoietic precursor cells, stage-specific analysis of T cell maturation confirmed early arrest of T cell differentiation at the T cell progenitor stage in X-SCID cells. In contrast, genetically corrected ESCs differentiated to CD4 + or CD8 + single-positive T cells, confirming correction of the cellular X-SCID phenotype. This study emphasises the value of PSCs for disease modelling and underlines the significance of in vitro models as tools to validate genome editing strategies before clinical application.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Substitution
  • Animals
  • Cell Differentiation
  • Disease Models, Animal
  • Gene Editing / methods*
  • Gene Expression
  • Hematopoietic Stem Cells / drug effects
  • Hematopoietic Stem Cells / immunology*
  • Hematopoietic Stem Cells / pathology
  • Humans
  • Interleukin Receptor Common gamma Subunit / deficiency
  • Interleukin Receptor Common gamma Subunit / genetics*
  • Interleukin Receptor Common gamma Subunit / immunology
  • Interleukin-2 / genetics
  • Interleukin-2 / immunology
  • Interleukin-2 / pharmacology
  • Interleukin-7 / genetics
  • Interleukin-7 / immunology
  • Interleukin-7 / pharmacology
  • Killer Cells, Natural / drug effects
  • Killer Cells, Natural / immunology
  • Killer Cells, Natural / pathology
  • Mice
  • Mice, SCID
  • Mice, Transgenic
  • Molecular Targeted Therapy
  • Mouse Embryonic Stem Cells / drug effects
  • Mouse Embryonic Stem Cells / immunology*
  • Mouse Embryonic Stem Cells / pathology
  • Mutation
  • T-Lymphocytes / drug effects
  • T-Lymphocytes / immunology
  • T-Lymphocytes / pathology
  • Transcription Activator-Like Effector Nucleases / genetics
  • Transcription Activator-Like Effector Nucleases / immunology
  • Transgenes
  • X-Linked Combined Immunodeficiency Diseases / genetics*
  • X-Linked Combined Immunodeficiency Diseases / immunology
  • X-Linked Combined Immunodeficiency Diseases / pathology
  • X-Linked Combined Immunodeficiency Diseases / therapy*


  • IL2RG protein, human
  • Interleukin Receptor Common gamma Subunit
  • Interleukin-2
  • Interleukin-7
  • interleukin-7, mouse
  • Transcription Activator-Like Effector Nucleases