The Effects of Daxx Knockout on Pluripotency and Differentiation of Mouse Induced Pluripotent Stem Cells

• Published in: Cellular reprogramming Vol. 22; no. 2; p. 90
• Main Authors: Liu, Hui, Liu, Zhaojun, Gao, Meng, Hu, Xinglin, Sun, Ruizhen, Shen, Xinghui, Liu, Feng, Shen, Jingling, Shan, Zhiyan, Lei, Lei
• Format: Journal Article
• Language: English
• Published: United States 01.04.2020
• Subjects: knockout; induced pluripotent stem cells; differentiation; Daxx
• ISSN: 2152-4998
• DOI: 10.1089/cell.2019.0071

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 mice, which carry the Oct4-green fluorescent protein reporter, and employed wild-type littermates as a control to induce iPSCs, then knocked out 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).