Overexpression of NEUROG2 and NEUROG1 (NEUROG2/1) in human embryonic stem cells (hESCs) rapidly produces functional networks of excitatory and inhibitory neurons. To facilitate the use of this efficient inducible human neuron model in neuroscience research, we generated hESCs with doxycycline-inducible NEUROG2/1 via lentivirus and a tdTomato fluorescent reporter knock-in at the MAP2 locus using the CRISPR nuclease Cas9. Upon doxycycline-driven induction of NEUROG2/1, these hESCs differentiate within days into cells that are uniformly MAP2 immunoreactive and tdTomato fluorescent.

中文翻译：

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生成具有 NEUROG2/1 (NYGCe001-A) 可诱导表达的敲入 MAP2-tdTomato 报告基因人胚胎干细胞系。

NEUROG2和NEUROG1 ( NEUROG2/1 ) 在人类胚胎干细胞 (hESCs) 中的过度表达会迅速产生兴奋性和抑制性神经元的功能网络。为了促进在神经科学研究中使用这种有效的诱导型人类神经元模型，我们通过慢病毒和 tdTomato 荧光报告基因敲入使用 CRISPR 核酸酶 Cas9在MAP2基因座上生成了具有强力霉素诱导型NEUROG2/1 的hESC 。在多西环素驱动的NEUROG2/1诱导后，这些 hESC 在数天内分化为具有一致 MAP2 免疫反应性和 tdTomato 荧光的细胞。