Modeling Human Severe Combined Immunodeficiency and Correction by CRISPR/Cas9-Enhanced Gene Targeting

Cell Rep. 2015 Sep 8;12(10):1668-77. doi: 10.1016/j.celrep.2015.08.013. Epub 2015 Aug 28.


Mutations of the Janus family kinase JAK3 gene cause severe combined immunodeficiency (SCID). JAK3 deficiency in humans is characterized by the absence of circulating T cells and natural killer (NK) cells with normal numbers of poorly functioning B cells (T(-)B(+)NK(-)). Using SCID patient-specific induced pluripotent stem cells (iPSCs) and a T cell in vitro differentiation system, we demonstrate a complete block in early T cell development of JAK3-deficient cells. Correction of the JAK3 mutation by CRISPR/Cas9-enhanced gene targeting restores normal T cell development, including the production of mature T cell populations with a broad T cell receptor (TCR) repertoire. Whole-genome sequencing of corrected cells demonstrates no CRISPR/Cas9 off-target modifications. These studies describe an approach for the study of human lymphopoiesis and provide a foundation for gene correction therapy in humans with immunodeficiencies.

MeSH terms

  • Bacterial Proteins / genetics
  • Base Sequence
  • CRISPR-Associated Protein 9
  • Cells, Cultured
  • Child, Preschool
  • Clustered Regularly Interspaced Short Palindromic Repeats
  • DNA Mutational Analysis
  • Endonucleases / genetics
  • Gene Targeting
  • Genetic Therapy*
  • Humans
  • Induced Pluripotent Stem Cells / enzymology
  • Janus Kinase 3 / genetics*
  • Male
  • Mutation, Missense
  • Proto-Oncogene Proteins c-bcl-2 / metabolism
  • Severe Combined Immunodeficiency / genetics
  • Severe Combined Immunodeficiency / therapy*
  • T-Lymphocytes / physiology


  • BCL2 protein, human
  • Bacterial Proteins
  • Proto-Oncogene Proteins c-bcl-2
  • JAK3 protein, human
  • Janus Kinase 3
  • CRISPR-Associated Protein 9
  • Cas9 protein, Francisella novicida
  • Endonucleases

Associated data

  • SRA/SRP056149