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

被引：97

作者：

Chang, Chia-Wei ^{[1
,5
,6
,10
]}

Lai, Yi-Shin ^{[1
,5
,6
,10
]}

Westin, Erik ^{[1
,5
,6
,10
]}

Khodadadi-Jamayran, Alireza ^{[1
,5
,6
,10
]}

Pawlik, Kevin M. ^{[1
,5
,6
,10
]}

Lamb, Lawrence S., Jr. ^{[2
,3
,5
,7
,8
,10
]}

Goldman, Frederick D. ^{[4
,5
,9
,10
]}

Townes, Tim M. ^{[1
,5
,6
,10
]}

机构：

[1] Univ Alabama Birmingham, Sch Med, Dept Biochem & Mol Genet, Birmingham, AL 35294 USA

[2] Univ Alabama Birmingham, Sch Med, Dept Med, Div Hematol Oncol, Birmingham, AL 35294 USA

[3] Univ Alabama Birmingham, Sch Med, Cell Therapy Lab, Birmingham, AL 35294 USA

[4] Univ Alabama Birmingham, Sch Med, Dept Pediat, Div Hematol Oncol, Birmingham, AL 35294 USA

[5] Univ Alabama Birmingham, Sch Med, UAB Stem Cell Inst, Birmingham, AL 35294 USA

[6] Univ Alabama Birmingham, Sch Dent, Dept Biochem & Mol Genet, Birmingham, AL 35294 USA

[7] Univ Alabama Birmingham, Sch Dent, Dept Med, Div Hematol Oncol, Birmingham, AL 35294 USA

[8] Univ Alabama Birmingham, Sch Dent, Cell Therapy Lab, Birmingham, AL 35294 USA

[9] Univ Alabama Birmingham, Sch Dent, Dept Pediat, Div Hematol Oncol, Birmingham, AL 35294 USA

[10] Univ Alabama Birmingham, Sch Dent, UAB Stem Cell Inst, Birmingham, AL 35294 USA

来源：

CELL REPORTS | 2015年 / 12卷 / 10期

关键词：

T-CELL DEVELOPMENT; PLURIPOTENT STEM-CELLS; MICE LACKING JAK3; IN-VITRO; LYMPHOID DEVELOPMENT; BCL-2; EXPRESSION; DEFICIENT MICE; GAMMA-CHAIN; PAIRED-END; INDUCTION;

D O I：

10.1016/j.celrep.2015.08.013

中图分类号：

Q2 [细胞生物学];

学科分类号：

071009 ; 090102 ;

摘要：

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.

引用

页码：1668 / 1677

页数：10

共 55 条

[21] Defective development of γ/δ T cells in interleukin 7 receptor-deficient mice is due to impaired expression of T cell receptor γ genes [J].