【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。更多还原

【Abstract】 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.更多还原