Methyladenosine modification of circRNA-ZNF638 contributes to the induced activation of SHF stem cells through miR-361-5p/Wnt5a axis in cashmere goats

Objective: The objective of this study was to investigate the effects of [N.sup.6]-Methyladenosine modification-circRNA-zinc finger protein 638 ([m.sup.6]A-circRNA-ZNF638) on the induced activation of secondary hair follicle (SHF) stem cells with its potential mechanisms in cashmere goats. Methods: The [m.sup.6]A modification of ZNF638 was analyzed using methylation immunoprecipitation with real-time quantitative polymerase chain reaction technique in SHF stem cells. The effects of circRNA-ZNF638 on the induced activation of SHF stem cells in [m.sup.6]A dependence were evaluated through the overexpression of circRNA-ZNF638/its [m.sup.6]A-deficient mutants in circRNA-ZNF638 knockdown SHF stem cells. The competitive binding of miR-361-5p to circRNA-ZNF638/Wnt5a 3'-untranslated region was analyzed through Dual-luciferase reporter assay. Results: The [m.sup.6]A-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, [m.sup.6]A-circRNA-ZNF638 sponged miR-361-5p to heighten the transcriptional expression of Wnt5a gene in SHF-stem cells. We further demonstrated that the internal [m.sup.6]A 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 [m.sup.6]A-deficient mutant of circRNA-ZNF638. Conclusion: The circRNA-ZNF638 contributes the proper induced activation of SHF-stem cells in cashmere goats in [m.sup.6]A-dependent manner through miR-361-5p/Wnt5a axis. Keywords: Cashmere Goats; CircRNA-ZNF638; MiR-361-5p; [N.sup.6]-Methyladenosine; SHF Stem Cells; Wnt5a