Induced pluripotent stem cell (iPSC) technology refers to the reprogramming of terminally differentiated somatic cells into pluripotent stem cells by introducing specific transcription factors that are known to regulate pluripotency, including Oct4, Sox2, Klf4, and c-Myc. In this study, we reprogrammed the primary fibroblasts isolated from the Daxx^flox/flox mice, which carry the Oct4-green fluorescent protein reporter, and employed wild-type littermates as a control to induce iPSCs, then knocked out Daxx by infecting with Cre virus at the cellular level. The pluripotency and self-renewal capacity of iPSCs were determined. In addition, Daxx deletion altered the pluripotency marker (Nanog, Oct4) expression and displayed neural differentiation defects. Particularly, by performing transcriptome analysis, we observed that numerous ribosome biogenesis-related genes were altered, and quantitative polymerase chain reaction revealed that the expression of rDNA-related genes, 47S and 18S, was elevated after Daxx deletion. Finally, we illustrated that the expression of the neurodevelopment-related gene was upregulated both in iPSCs and differentiated neurospheres. Taken together, we demonstrated that Daxx knockout promotes the expression of rDNA, pluripotency, and neurodevelopment genes, which may improve the differentiation abilities of mouse iPSCs (miPSCs).

中文翻译：

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Daxx基因敲除对小鼠诱导的多能干细胞多能性和分化的影响。

诱导多能干细胞（iPSC）技术是指通过引入已知可调节多能性的特定转录因子（包括Oct4，Sox2，Klf4和c-Myc）将最终分化的体细胞重编程为多能干细胞。在这项研究中，我们对从Daxx ^{flox / flox}小鼠中分离的原代成纤维细胞进行了重新编程，这些小鼠带有^Oct4-绿色荧光蛋白报告基因，并使用野生型同窝幼虫作为诱导iPSC的对照，然后敲除了Daxx通过在细胞水平感染Cre病毒。确定了iPSC的多能性和自我更新能力。此外，Daxx缺失改变了多能性标记（Nanog，Oct4）的表达并显示出神经分化缺陷。特别地，通过进行转录组分析，我们观察到许多核糖体生物发生相关基因被改变，并且定量聚合酶链反应显示在Daxx缺失后rDNA相关基因47S和18S的表达升高。最后，我们说明了神经发育相关基因的表达在iPSC和分化的神经球中均被上调。综上所述，我们证明了Daxx基因敲除可促进rDNA的表达，多能性和神经发育基因，