生成三个具有 DOX 诱导 NGN2 表达盒的同源基因编辑亨廷顿病人类胚胎干细胞系，位于 AAVS1 安全位点。
Generation of three isogenic gene-edited Huntington's disease human embryonic stem cell lines with DOX-inducible NGN2 expression cassette in the AAVS1 safe locus.
机构信息
Dept. of Cellular and Molecular Medicine, The Panum Institute, The Faculty of Health and Medical Sciences, University of Copenhagen, Copenhagen 2200N, Denmark.
Department of Veterinary and Animal Sciences, Faculty of Health and Medical Sciences, University of Copenhagen, 1870 Frederiksberg, Denmark.
出版信息
Stem Cell Res. 2024 Jun;77:103408. doi: 10.1016/j.scr.2024.103408. Epub 2024 Mar 28.
Neurogenin 2 (NGN2), a neuronal transcription factor, can expedite differentiation of stem cells into mature glutamatergic neurons. We have utilized an allelic series of previously published and characterized isogenic Huntington's disease (IsoHD) human embryonic stem cell lines (Ooi et al., 2019), carrying different CAG repeat lengths in the first exon of the huntingtin gene. These IsoHDs were modified using CRISPR/Cas9 to insert NGN2 under the TET-ON doxycycline inducible promoter. The resulting IsoHD-NGN2 cell lines retained pluripotency in the absence of doxycycline (DOX), and via addition of DOX to the culturing media differentiation to neurons was achieved within 14 days.
神经基因 2（NGN2）是一种神经元转录因子，能够加速干细胞向成熟谷氨酸能神经元的分化。我们利用了一系列先前发表并具有特征的同型亨廷顿病（IsoHD）人类胚胎干细胞系（Ooi 等人，2019 年），这些细胞系在亨廷顿基因的第一个外显子中携带不同的 CAG 重复长度。这些 IsoHD 通过 CRISPR/Cas9 进行修饰，将 NGN2 插入 Tet-On 诱导型多西环素启动子下。在没有多西环素（DOX）的情况下，所得的 IsoHD-NGN2 细胞系保留了多能性，并且通过在培养介质中添加 DOX，在 14 天内实现了向神经元的分化。
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