MicroRNA Cluster 290-295 Enhances Somatic Cell Reprogramming

doi:10.13523/j.cb.2211006

China Biotechnology

2023, Vol. 43

Issue (4): 1-9 DOI: 10.13523/j.cb.2211006

MicroRNA Cluster 290-295 Enhances Somatic Cell Reprogramming

ZHU Si-ying^1,²,YANG Yang²,LI Peng-dong²,XUE Yan-ting³,SHE Qin²,QI Ling²,ZHAO Guo-jun^2,^**(

),LIAO Bao-jian^2,^3,^**(

)

1. College of Pharmacy, Dali University, Dali 671000, China
2. The Sixth Affiliated Hospital of Guangzhou Medical University, Qingyuan 511518, China
3. Guangzhou Institutes of Biomedicine and Health, Chinese Academy of Sciences, Guangzhou 510530, China

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Abstract

Objective: To explore the effect of miR-290-295, the most abundantly expressed microRNA cluster in stem cells, as a whole on somatic cell reprogramming. Methods: The miR-290-295 cluster was overexpressed into mouse somatic cells using retroviral vectors to explore whether it promotes the reprogramming of somatic cells into induced pluripotent stem cells (iPSCs) and the effect on cell function through this process. Results: The overexpression of miR-290-295 cluster could significantly improve the efficiency of mouse somatic cell reprogramming in the traditional induction system of three factors (Sox2, Klf4 and Oct4). Overexpression of miR-290-295 cluster could not only promote the up-regulation of pluripotency marker genes and the down-regulation of somatic marker genes during reprogramming, but also the expression of mesenchymal-epithelial transition (MET) marker genes and cell proliferation related genes. Conclusion: miR-290-295 promotes the reprogramming of mouse somatic cells. Our findings are helpful to understand the RNA regulatory mechanism in stem cell pluripotency and reprogramming, and provide a new perspective for the development of new induction systems.

Key words： Somatic cell reprogramming Induced pluripotent stem cells MicroRNA MiR-290-295 cluster

Received: 03 November 2022 Published: 04 May 2023

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	Si-ying ZHU
	Yang YANG
	Peng-dong LI
	Yan-ting XUE
	Qin SHE
	Ling QI
	Guo-jun ZHAO
	Bao-jian LIAO

Cite this article:

ZHU Si-ying, YANG Yang, LI Peng-dong, XUE Yan-ting, SHE Qin, QI Ling, ZHAO Guo-jun, LIAO Bao-jian. MicroRNA Cluster 290-295 Enhances Somatic Cell Reprogramming. China Biotechnology, 2023, 43(4): 1-9.

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https://manu60.magtech.com.cn/biotech/10.13523/j.cb.2211006 OR https://manu60.magtech.com.cn/biotech/Y2023/V43/I4/1

Fig.1 Efficiency of overexpression of miR-290-295 cluster (a) MiRNA profiling in mESCs and Plat-E by small RNA sequencing (b) Principles of RT-qPCR detection of miRNAs (c) Efficiency detection of overexpressed miR-290-295 clusters. Compared with flag control, ^*** indicate P<0.001, highly significant, n=3

Table 1 Primers for RT-qPCR

Fig.2 Overexpression of miR-290-295 cluster promotes reprogramming (a) Efficiency detection of overexpressed miR-290-295 clusters during reprogramming, n=3 (b)(c) Comparison of the number of Oct4-GFP positive clones after overexpression of miR-290-295 cluster. Compared with flag control, ^*** indicate P<0.001, highly significant, n=3 (d) RT-qPCR analysis for the pluripotency marker genes, n=3 (e) Immunofluorescent staining of generated iPSCs

Fig.3 miR-290-295 cluster promotes reprogramming through classical MET transition (a) Effect of overexpression of miR-290-295 on the expression of pluripotent marker genes (b) Effect of overexpression of miR-290-295 on the expression of mesenchymal-epithelial transition marker genes (Snai1, Snai2,Twist2, Cdh2 are mesenchymal cell marker genes, Cdh1, Epcam are epithelial marker genes), n=3


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