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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
Stem Cell Research ( IF 1.2 ) Pub Date : 2024-03-28 , DOI: 10.1016/j.scr.2024.103408
Luisana Duque Villegas , Abinaya Chandrasekaran , Sofie Amalie Flintholm Andersen , Anne Nørremølle , Benjamin Schmid , Mahmoud A. Pouladi , Kristine Freude

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 (), carrying different CAG repeat lengths in the first exon of the huntingtin gene. These IsoHDs were modified using CRISPR/Cas9 to insert under the TET-ON doxycycline inducible promoter. The resulting IsoHD- 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.

中文翻译:

生成三种等基因基因编辑的亨廷顿病人胚胎干细胞系,其 AAVS1 安全位点具有 DOX 诱导的 NGN2 表达盒

Neurogenin 2 (NGN2) 是一种神经元转录因子,可以加速干细胞分化为成熟的谷氨酸能神经元。我们利用了一系列先前发表和表征的同基因亨廷顿病 (IsoHD) 人类胚胎干细胞系 (),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 使用 CRISPR/Cas9 进行修饰,插入到 TET-ON 多西环素诱导型启动子下。所得的 IsoHD 细胞系在没有强力霉素 (DOX) 的情况下保留了多能性,并且通过向培养基中添加 DOX,在 14 天内实现了向神经元的分化。
更新日期:2024-03-28
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