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https://doi.org/10.5713/ab.22.0211    [Accepted] Published online November 14, 2022.
N6-Methyladenosine modification (m6A) of circRNA-ZNF638 contributes the induced activation of SHF stem cells through miR-361-5p/Wnt5a axis in cashmere goats
Ronghuan Yin1  , Ronglan Yin2  , Man Bai1  , Yixing Fan1  , Zeying Wang1  , Yubo Zhu1  , Qi Zhang1  , Taiyu Hui1  , Jincheng Shen1  , Siyu Feng1  , Wenlin Bai1,* 
1College of Animal Science & Veterinary Medicine, Shenyang Agricultural University, Shenyang 110866, China
2Research Academy of Animal Husbandry and Veterinary Medicine Sciences of Jilin Province, Changchun, 130062, China
Correspondence:  Wenlin Bai, Tel: +86-24-88487156, Fax: +86-24-88487156, Email: baiwenlin@syau.edu.cn
Received: 27 May 2022   • Revised: 30 July 2022   • Accepted: 1 October 2022
Abstract
Objective
The objective of this study was to investigate the effects of N6-Methyladenosine modification-circRNA-zinc finger protein 638 (m6A-circRNA-ZNF638) on the induced activation of secondary hair follicle (SHF) stem cells with its potential mechanisms in cashmere goats.
Methods
The m6A modification of ZNF638 was analyzed using Methylation Immunoprecipitation (Me-RIP) with real-time quantitative PCR (RT-qPCR) technique in SHF stem cells. The effects of circRNA-ZNF638 on the induced activation of SHF stem cells in m6A dependence were evaluated through the overexpression of circRNA-ZNF638/its m6A-deficient mutants in circRNA-ZNF638 knockdown SHF stem cells. The competitive binding of miR-361-5p to circRNA-ZNF638/Wnt5a 3′- untranslated region (3′-UTR) was analyzed through Dual-luciferase reporter assay.
Results
The m6A-circRNA-ZNF638 had significantly higher transcription at anagen SHF bulge of cashmere goats compared with that at telogen, as well as it positively regulated the induced activation of SHF-stem cells in cashmere goats. Mechanismly, m6A-circRNA-ZNF638 sponged miR-361-5p to heighten the transcriptional expression of Wnt5a gene in SHF-stem cells. We further demonstrated that the internal m6A modification within circRNA-ZNF638 is required for mediating the miR-361-5p/Wnt5a pathway to regulate the induced activation of SHF stem cells through an introducing of m6A-deficient mutant of circRNA-ZNF638.
Conclusion
The circRNA-ZNF638 contributes the proper induced activation of SHF-stem cells in cashmere goats in m6A-dependent manner through miR-361-5p/Wnt5a axis.
Keywords: Cashmere Goats; CircRNA-ZNF638; MiR-361-5p; N6-Methyladenosine; SHF Stem Cells; Wnt5a
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