The generation and validation of two NKX2-5 fluorescent reporter human embryonic stem cell lines: UMANe002-A-1 and UMANe002-A-2
The generation and validation of two NKX2-5 fluorescent reporter human embryonic stem cell lines: UMANe002-A-1 and UMANe002-A-2
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两种 NKX2-5 荧光报告基因人胚胎干细胞系 UMANe002-A-1 和 UMANe002-A-2 的生成和验证
DOI：
10.1016/j.scr.2023.103262
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发表时间：
2024
影响因子：
1.2
通讯作者：
Douglas M
中科院分区：
文献类型：
--
作者：
Douglas M
转录因子NKX 2 -5是心脏发育的高度保守的主调节因子，其在心脏祖细胞和心肌细胞中广泛表达。NKX 2 - 5的荧光报告基因对正常蛋白质表达的干扰最小，可以在正常生理状态下识别、定量和分离表达NKX 2 -5的细胞。在此我们报道了两个新的hESC细胞系的产生，GFP插入NKX 2 -5的上游（5′）或下游（3′），通过可切割的T2 A肽连接。这些互补报告基因在心脏细胞中产生强大的荧光信号，特别是在发育生物学和疾病建模研究中具有广泛的用途。
The transcription factor NKX2-5 is a highly conserved master regulator of heart development which is widely expressed in cardiac progenitors and cardiomyocytes. Fluorescent reporters ofNKX2-5that minimally perturb normal protein expression can enable the identification, quantification and isolation of NKX2-5-expressing cells in a normal physiological state. Here we report the generation of two new hESC lines witheGFPinserted upstream (5′) or downstream (3′) ofNKX2-5, linked by a cleavable T2A peptide. These complementary reporters produce a robust fluorescent signal in cardiac cells and have wide utility particularly for research on developmental biology and disease modelling.