Ramovs Veronika, Fuentes Ignacia, Freund Christian, Mikkers Harald, Mummery Christine L, Raymond Karine
Department of Anatomy and Embryology, Leiden University Medical Center, Leiden 2333 ZC, The Netherlands.
Fundación DEBRA Chile, Santiago, Chile; Centro de Genética y Genómica, Facultad de Medicina, Clínica Alemana Universidad del Desarrollo, Santiago, Chile.
Stem Cell Res. 2021 Dec;57:102582. doi: 10.1016/j.scr.2021.102582. Epub 2021 Oct 21.
Fibroblasts from two patients carrying a heterozygous mutation in the translation initiation codon (c.2 T > G) of the kelch-like protein 24 (KLHL24) gene were used to generate human induced pluripotent stem cells (hiPSCs), using non-integrating Sendai virus to deliver reprogramming factors. CRISPR-Cas9 editing was used for genetic correction of the mutation in the patient-hiPSCs. The top-predicted off-target sites were not altered. Patient and isogenic hiPSCs showed typical morphology, expressed pluripotency-associated markers, had the capacity for in vitro differentiation into the three germ layers and displayed a normal karyotype. These isogenic pairs will enable in vitro modelling of KLHL24-associated heart and skin conditions.
利用携带kelch样蛋白24(KLHL24)基因翻译起始密码子杂合突变(c.2 T>G)的两名患者的成纤维细胞,通过非整合仙台病毒递送重编程因子来生成人类诱导多能干细胞(hiPSC)。CRISPR-Cas9编辑用于对患者来源的hiPSC中的突变进行基因校正。预测的脱靶位点没有改变。患者来源的hiPSC和同基因hiPSC表现出典型的形态,表达多能性相关标志物,具有体外分化为三个胚层的能力,并且核型正常。这些同基因细胞对将有助于对与KLHL24相关的心脏和皮肤疾病进行体外建模。