Hao Engang, Zhang Guizhong, Mu Lihua, Ma Nana, Wang Tao
Department of Cardiology, The Second People's Hospital of Liaocheng, Linqing 252600, China.
Department of Cardiology, The Second People's Hospital of Liaocheng, Linqing 252600, China.
Stem Cell Res. 2021 Jan;50:102128. doi: 10.1016/j.scr.2020.102128. Epub 2020 Dec 16.
MYH6 encodes the alpha heavy chain subunit of cardiac myosin. Mutations in MYH6 cause cardiomyopathy and congenital heart defects. However, due to embryonic lethality in MYH6 knockout mice, the precise roles of MYH6 in cardiomyopathy, congenital heart defects and development process remain largely unknown. In this study, we generated a human MYH6 compound heterozygous knockout hESC line using CRISPR/Cas9 technology. The establishment cell line WAe009-A-46 carried a compound heterozygous 2 bp deletion/7 bp deletion in MYH6, expressed pluripotency markers, showed a normal karyotype and exhibited capability to differentiate into the three germ layers in vitro. MYH6 protein was not detectable in WAe009-A-46 line. This cell line provides a useful tool for studying the role of MYH6 in cardiomyopathy and congenital heart defects.
MYH6编码心肌肌球蛋白的α重链亚基。MYH6突变会导致心肌病和先天性心脏缺陷。然而，由于MYH6基因敲除小鼠存在胚胎致死性，MYH6在心肌病、先天性心脏缺陷和发育过程中的精确作用在很大程度上仍不清楚。在本研究中，我们使用CRISPR/Cas9技术构建了一株人MYH6复合杂合敲除的人胚胎干细胞系。建立的细胞系WAe009-A-46在MYH6中存在复合杂合的2bp缺失/7bp缺失，表达多能性标志物，核型正常，且在体外具有分化为三个胚层的能力。在WAe009-A-46细胞系中未检测到MYH6蛋白。该细胞系为研究MYH6在心肌病和先天性心脏缺陷中的作用提供了一个有用的工具。
