基于基因编辑技术的DMD人诱导多能干细胞模型构建  

Construction of DMD hiPSC Cell Lines by CRISPR/Cas9 Editing

作　　者：王伟[1] 林翔[1] 陈万金[1] 王柠[1] 金铭 WANG Wei;LIN Xiang;CHEN Wanjin;WANG Ning;JIN Ming(Department of Neurology,The First Affiliated Hospital of Fujian Medical University,Fuzhou 350005,China)

机构地区：[1]福建医科大学附属第一医院神经内科,福州350005

出　　处：《福建医科大学学报》2021年第3期181-186,共6页Journal of Fujian Medical University

基　　金：国家自然科学基金面上项目(81870902);福建省科技创新联合资金项目(2018Y9082)。

摘　　要：目的采用CRISPR/Cas9技术,构建杜氏进行性肌营养不良(DMD)大片段缺失的人诱导多能干细胞(hiPSCs)模型。方法采用电转质粒的方式,在正常的hiPSCs上用双sgRNA切割方案,切除dystrophin基因的52号外显子。结果T7E1分析显示,所选sgRNA切割效率最高可达91%,后续基因型鉴定显示成功获得了52号外显子切除的hiPSCs细胞系。结论基因编辑构建的hiPSCs模型可以作为后续DMD基因编辑治疗研究的体外细胞模型。Objective To construct hiPSCs cell lines of Duchenne muscular dystroply by CRISPR/Cas9 editing technology. Methods The 52 nd exon of dystrophin gene was excised by duel-sgRNA strategy using nucleofector. Result T7 E1 assay showed the cleavage efficiency could reach 91%. Subsequent genotyping analysis indicated that in some cell lines the 52 nd exon of dystrophin had been excised. Conclusion The hiPSCs of dystrophin can be used for research as cell line models in vitro.

关 键 词：杜氏进行性肌营养不良 人诱导多能干细胞 基因编辑 模型构建 

分 类 号：R746.2[医药卫生—神经病学与精神病学]