利用基于 CRISPR/Cas9 的基因靶向技术，在人胚胎干细胞系 WAe001-A-2E 中生成 ASCL1-mCherry 敲入报告基因。
Generation of ASCL1-mCherry knock-in reporter in human embryonic stem cell line, WAe001-A-2E, using CRISPR/Cas9-based gene targeting.
机构信息
School of Biomedical Engineering, Guangzhou Medical University, Guangzhou 510180, China; Guangzhou National Laboratory, Guangzhou 510005, China.
College of Life Science and Technology, Huazhong University of Science and Technology, Wuhan 430074, China; Guangzhou National Laboratory, Guangzhou 510005, China.
出版信息
Stem Cell Res. 2024 Oct;80:103500. doi: 10.1016/j.scr.2024.103500. Epub 2024 Jul 24.
Achaete-Scute Complex Homolog 1 (ASCL1) is a key regulator in the development and function of the nervous system, particularly in the process of neuronal and neuroendocrine cell differentiation. By employing the CRISPR/Cas9 system, we successfully established an ASCL1-mCherry knock-in human embryonic stem cell (hESC) line by inserting a P2A-mCherry fragment at the ASCL1 locus. The mCherry reporter effectively demonstrated the expression level of endogenous ASCL1 during the process of inducing pulmonary neuroendocrine cells (PNECs) from hESC. This reporter cell line holds significant value as a research tool for investigating the process of lung neuroendocrine cell differentiation, conducting drug screening, and exploring the underlying mechanisms of lung diseases associated with PNECs dysfunction.
achaete-scute 复合体同源物 1（ASCL1）是神经系统发育和功能的关键调节因子，特别是在神经元和神经内分泌细胞分化的过程中。我们使用 CRISPR/Cas9 系统，成功地在 ASCL1 基因座插入一个 P2A-mCherry 片段，建立了一个 ASCL1-mCherry 敲入的人胚胎干细胞（hESC）系。mCherry 报告基因在诱导 hESC 产生肺神经内分泌细胞（PNEC）的过程中，有效地显示了内源性 ASCL1 的表达水平。该报告细胞系作为研究工具具有重要价值，可用于研究肺神经内分泌细胞分化的过程、进行药物筛选以及探索与 PNEC 功能障碍相关的肺部疾病的潜在机制。
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