Institute of Reproductive Medicine and Population, Medical Research Center, Dept. of Obstetrics & Gynecology, Seoul National University College of Medicine, Seoul 03080, Republic of Korea.
Department of Biochemistry, Institute for Aging and Metabolic Diseases, College of Medicine, The Catholic University of Korea, 222, Banpo-daero, Seocho-gu, Seoul 06591, Republic of Korea.
Stem Cell Res. 2021 May;53:102321. doi: 10.1016/j.scr.2021.102321. Epub 2021 Apr 10.
Brachyury is an embryonic nuclear transcription factor required for mesoderm formation and differentiation. Here, we introduced an mCherry reporter into the C-terminus of Brachyury in the human pluripotent stem cell line SNUhES3 using the CRISPR/Cas9 nuclease approach. Successful gene editing was verified by DNA sequencing. SNUhES3-Brachyury-mCherry cells expressed pluripotent stem cell markers, exhibited a normal karyotype, and could generate all three germ layers. This cell line expressed the red fluorescence protein mCherry upon the induction of mesoderm differentiation. This reporter cell line could be used to monitor mesodermal population enrichment during mesodermal differentiation.
Brachyury 是一种胚胎核转录因子，对于中胚层的形成和分化是必需的。在这里，我们使用 CRISPR/Cas9 核酸酶方法将 mCherry 报告基因插入人多能干细胞系 SNUhES3 中 Brachyury 的 C 末端。通过 DNA 测序验证了成功的基因编辑。SNUhES3-Brachyury-mCherry 细胞表达多能干细胞标志物，具有正常的核型，并能产生所有三个胚层。在诱导中胚层分化时，该细胞系表达红色荧光蛋白 mCherry。该报告细胞系可用于监测中胚层分化过程中中胚层群体的富集。
