Genome wide gene-expression and mRNA splicing profile in human cardiomyocytes differentiated from Wt and QKI mutant embryonic stem cells

We sequenced mRNA from cardiomyocytes derived from hESCS in vitro. By using the Cas9n, we generated the QKI null hESCs. The gene expression level and alternative splicing events were compared between 4 control and 4 QkI KO samples. Here, we applied a widely used cardiomyocyte differentiation protocol that was reported to produce a population of more than 90% cardiac troponin T (TNNT2)-positive cardiomyocytes. And we are able to demonstrate that QKI is indespensible to cardiac sarcomerogenesis and cardiac function through its regulation of alternative splicing in genes involved in Z-disc formation, such as ACTN2, NEBL, ABLIM1, and PDLIM5. Overall design: Compare potential altered RNA processing in normal and QKI mutant hESCs-differentiated hCMs at D15 of differention and alternative RNA splicing changes.

我们对体外（in vitro）由人类胚胎干细胞（human embryonic stem cells, hESCS）分化得到的心肌细胞进行了mRNA测序。借助Cas9切口酶（Cas9n），我们构建了QKI基因完全敲除的人类胚胎干细胞。本研究对4组对照样本与4组QKI敲除样本的基因表达水平及可变剪接事件开展了对比分析。本研究采用了一项已被报道可获得超过90%心肌肌钙蛋白T（cardiac troponin T, TNNT2）阳性心肌细胞的通用心肌细胞分化方案。研究证实，QKI可通过调控参与Z盘形成的基因（如ACTN2、NEBL、ABLIM1及PDLIM5）的可变剪接过程，对心肌肌节发生及心脏功能发挥不可或缺的作用。实验设计概述：比较正常及QKI突变人类胚胎干细胞分化得到的心肌细胞在分化第15天时的潜在RNA加工改变与可变剪接变化。