Modeling Human Severe Combined Immunodeficiency and Correction by CRISPR/Cas9-Enhanced Gene Targeting
Modeling Human Severe Combined Immunodeficiency and Correction by CRISPR/Cas9-Enhanced Gene Targeting
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DOI：
10.1016/j.celrep.2015.08.013
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发表时间：
2015-09-08
期刊：
影响因子：
8.8
通讯作者：
Townes, Tim M.
中科院分区：
文献类型：
--
作者：
Chang, Chia-Wei;Lai, Yi-Shin;Townes, Tim M.
Janus家族激酶JAK 3基因突变导致严重联合免疫缺陷（SCID）。人中JAK 3缺乏的特征在于缺乏循环T细胞和自然杀伤（NK）细胞以及正常数量的功能不良的B细胞（T-B+NK-）。使用SCID患者特异性诱导多能干细胞（iPSC）和T细胞体外分化系统，我们证明了JAK 3缺陷细胞的早期T细胞发育完全阻断。通过CRISPR/Cas9增强的基因靶向纠正JAK 3突变恢复了正常的T细胞发育，包括产生具有广泛T细胞受体（TCR）库的成熟T细胞群。校正细胞的全基因组测序证明没有CRISPR/ Cas9脱靶修饰。这些研究描述了人类淋巴细胞生成的研究方法，并为免疫缺陷患者的基因校正治疗提供了基础。
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.