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. Circular RNAs (circRNAs) have stable structures,being a covalently closed loop without 5′ and 3′ free ends.They can function as “miRNA sponges” in regulating the expression of theirtarget genes. It was thought that circRNAs are involved in the developmentof the secondary hair follicle (SHF) in cashmere goats. In our previousinvestigation, a new circRNA named circRNA-0100 was identified from theSHF of cashmere goats, but its function is unknown. In this work, we foundthat circRNA-0100 exhibited significantly higher expression at anagen SHFbulge than its counterpart at telogen in cashmere goats. Based on the use ofboth overexpression and siRNA interference assays, our data indicated thatcircRNA-0100 promoted the differentiation of cashmere goat SHF stem cells(SHF-SCs) into hair follicle lineage, which was evaluated by analyzing thetranscriptional level changes of six indicator genes in SHF-SCs of cashmeregoats. Using the RNA pull-down technique, we showed thatcircRNA-0100 served as “molecular sponges” of miR-153-3p in SHF-SCs.Through the use of dual-luciferase reporter assays, our data indicated thatcircRNA-0100 positively regulated the transcriptional expression of the KLF5gene via the miR-153-3p-mediated pathway. Ultimately, we showed thatcircRNA-0100 promoted the differentiation of SHF-SCs into hair lineage, whichmight be achieved via sequestering miR-153-3p to heighten the KLF5expression in SHF-SCs of cashmere goats. Our results provide novelscientific evidence for revealing the potential molecular regulatorymechanisms on the differentiation of SHF-SCs into hair lineage in cashmeregoats.

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CircRNA-0100 positively regulates the differentiation of cashmere goat SHF-SCs into hair follicle lineage via sequestering miR-153-3p to heighten the KLF5 expression