title: Modeling Human Severe Combined Immunodeficiency and Correction by CRISPR/Cas9-Enhanced Gene Targeting.
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.
authors: 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.
authors: Chia-Wei
authors: CW
authors: Chang
authors: 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.
authors: Yi-Shin
authors: YS
authors: Lai
authors: 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.
authors: Erik
authors: E
authors: Westin
authors: 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.
authors: Alireza
authors: A
authors: Khodadadi-Jamayran
authors: 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.
authors: Kevin M
authors: KM
authors: Pawlik
authors: 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.
authors: Lawrence S
authors: LS
authors: Lamb
authors: 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.
authors: Frederick D
authors: FD
authors: Goldman
authors: 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.
authors: Tim M
authors: TM
authors: Townes
mesh_terms: D001426:Bacterial Proteins / Q000235:genetics; D001483:Base Sequence; D000076987:CRISPR-Associated Protein 9; D002478:Cells, Cultured; D002675:Child, Preschool; D064112:Clustered Regularly Interspaced Short Palindromic Repeats; D004252:DNA Mutational Analysis; D004720:Endonucleases / Q000235:genetics; D018390:Gene Targeting; D015316:Genetic Therapy; D006801:Humans; D057026:Induced Pluripotent Stem Cells / Q000201:enzymology; D053616:Janus Kinase 3 / Q000235:genetics*; D008297:Male; D020125:Mutation, Missense; D019253:Proto-Oncogene Proteins c-bcl-2 / Q000378:metabolism; D016511:Severe Combined Immunodeficiency / Q000235:genetics / Q000628:therapy*; D013601:T-Lymphocytes / Q000502:physiology
chemical_list: C000595707:BCL2 protein, human; D001426:Bacterial Proteins; D019253:Proto-Oncogene Proteins c-bcl-2; C507933:JAK3 protein, human; D053616:Janus Kinase 3; D000076987:CRISPR-Associated Protein 9; C000600049:Cas9 protein, Francisella novicida; D004720:Endonucleases