A novel CRISPR-Cas9 strategy to target DYSTROPHIN mutations downstream of exon 44 in patient-specific DMD iPSCs

Here, we performed CRISPR-cas9 mediated genetic correction in DMD patient derived iPSC and then differentiated them into in vitro myotubes. Pairing bulk RNA sequencing between Wild-type (WT), DMD and the corrected counterparts we were able to differentiate the molecular changes that occurs during the absence of dystrophin. We performed gene expression profiling for data obtained through bulk RNA-sequencing of 15 uniques samples. Samples included WT, DMD and DMD corrected iPSC derived myotubes.

本研究针对杜氏肌营养不良症（Duchenne Muscular Dystrophy, DMD）患者来源的诱导多能干细胞（induced pluripotent stem cell, iPSC）实施CRISPR-Cas9（成簇规律间隔短回文重复序列相关蛋白9）介导的基因校正操作，随后将校正后的细胞定向分化为体外肌管。通过比对野生型（Wild-type, WT）、DMD模型及校正后对应样本的批量RNA测序（bulk RNA-sequencing）数据，我们得以清晰阐明抗肌萎缩蛋白（dystrophin）缺失过程中发生的分子改变。本研究对15个独立样本的批量RNA测序数据开展基因表达谱分析，所涉样本涵盖野生型、DMD模型及经校正的iPSC来源肌管。