Restoring Dystrophin Expression with Duchenne Muscular Dystrophy Exon 45 Skipping in Induced Pluripotent Stem Cell-Derived Cardiomyocytes
Restoring Dystrophin Expression with Duchenne Muscular Dystrophy Exon 45 Skipping in Induced Pluripotent Stem Cell-Derived Cardiomyocytes
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通过杜氏肌营养不良症外显子 45 跳跃恢复诱导多能干细胞来源的心肌细胞中肌营养不良蛋白的表达
DOI：
10.1007/978-1-0716-2772-3_8
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发表时间：
2023
影响因子：
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通讯作者：
Akinori Nakamura
中科院分区：
文献类型：
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作者：
Mitsuto Sato;Naoko Shiba;Daigo Miyazaki;Yuji Shiba;Akinori Nakamura
基于诱导多能干细胞（iPSC）的疾病模型是一种有用的工具，可以代表由于侵入性而无法接近的患者器官的病理生理学。在这里，我们提出了一种方法，以诱导分化的杜氏肌营养不良症（DMD）患者来源的iPSC成心肌细胞和恢复肌营养不良蛋白的表达外显子跳跃使用反义核酸。这涉及20天的多步培养过程，用于分化为心肌细胞，然后进行外显子跳跃实验。此外，RT-PCR，蛋白质印迹和免疫细胞化学用于确认肌营养不良蛋白表达的恢复。
Induced pluripotent stem cell (iPSC)-based disease model is a useful tool that can represent the pathophysiology of patient organs that are inaccessible due to invasiveness. Here, we present a method to induce differentiation of Duchenne muscular dystrophy (DMD) patient-derived iPSCs into cardiomyocytes and restore dystrophin expression by exon skipping using antisense nucleic acids. This involves a 20-day multi-step culture process for differentiation to cardiomyocytes, followed by exon-skipping experiments. Additionally, RT-PCR, western blotting, and immunocytochemistry are used to confirm the restoration of dystrophin expression.