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

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

Author information

1
Department of Biochemistry and Molecular Genetics, Schools of Medicine and Dentistry, University of Alabama at Birmingham, Birmingham, AL 35294, USA; UAB Stem Cell Institute, Schools of Medicine and Dentistry, University of Alabama at Birmingham, Birmingham, AL 35294, USA.
2
Department of Medicine, Division of Hematology/Oncology, Schools of Medicine and Dentistry, University of Alabama at Birmingham, Birmingham, AL 35294, USA; Cell Therapy Lab, Schools of Medicine and Dentistry, University of Alabama at Birmingham, Birmingham, AL 35294, USA; UAB Stem Cell Institute, Schools of Medicine and Dentistry, University of Alabama at Birmingham, Birmingham, AL 35294, USA.
3
Department of Pediatrics, Division of Hematology/Oncology, Schools of Medicine and Dentistry, University of Alabama at Birmingham, Birmingham, AL 35294, USA; UAB Stem Cell Institute, Schools of Medicine and Dentistry, University of Alabama at Birmingham, Birmingham, AL 35294, USA.
4
Department of Biochemistry and Molecular Genetics, Schools of Medicine and Dentistry, University of Alabama at Birmingham, Birmingham, AL 35294, USA; UAB Stem Cell Institute, Schools of Medicine and Dentistry, University of Alabama at Birmingham, Birmingham, AL 35294, USA. Electronic address: ttownes@uab.edu.

Abstract

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.

PMID:
26321643
DOI:
10.1016/j.celrep.2015.08.013
[Indexed for MEDLINE]
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