The Effects of Daxx Knockout on Pluripotency and Differentiation of Mouse Induced Pluripotent Stem Cells.
The Effects of Daxx Knockout on Pluripotency and Differentiation of Mouse Induced Pluripotent Stem Cells.
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Daxx 敲除对小鼠诱导多能干细胞的多能性和分化的影响。
DOI：
10.1089/cell.2019.0071
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发表时间：
2020-03
影响因子：
1.6
通讯作者：
Lei Lei
中科院分区：
文献类型：
--
作者：
Liu Hui;Liu Zhaojun;Gao Meng;Hu Xinglin;Sun Ruizhen;Shen Xinghui;Liu Feng;Shen Jingling;Shan Zhiyan;Lei Lei
诱导多能干细胞(IPSC)技术是指通过引入已知调节多能性的特定转录因子，包括Oct4、Sox2、Klf4和c-Myc，将终末分化的体细胞重新编程为多能干细胞。在本研究中，我们对携带Oct4绿色荧光蛋白报告基因的Daxxflx/FLOX小鼠的原代成纤维细胞进行了重编程，并以野生型窝种作为对照来诱导IPSCs，然后在细胞水平上通过感染Cre病毒来敲除Daxx。检测IPSCs的多能性和自我更新能力。此外，Daxx基因缺失改变了多能性标记物(Nanog，Oct4)的表达，并显示出神经分化缺陷。特别是，通过转录组分析，我们观察到许多与核糖体生物发生相关的基因发生了变化，定量聚合酶链式反应显示，在Daxx缺失后，rDNA相关基因47S和18S的表达上调。最后，我们阐明了神经发育相关基因在IPSCs和分化神经球中的表达上调。综上所述，我们证明Daxx基因敲除促进了rDNA、多能性和神经发育基因的表达，从而可能提高小鼠iPSCs的分化能力。
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 Daxxflox/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).
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影响因子：
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作者：
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DOI：
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发表时间：
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