Zhao Junyin, Shen Jincheng, Wang Zeying, Bai Man, Fan Yixing, Zhu Yubo, Bai Wenlin
College of Animal Science & Veterinary Medicine, Shenyang Agricultural University, Shenyang, 110866, P.R. China.
Arch Anim Breed. 2022 Feb 22;65(1):55-67. doi: 10.5194/aab-65-55-2022. eCollection 2022.
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 their target genes. It was thought that circRNAs are involved in the development of the secondary hair follicle (SHF) in cashmere goats. In our previous investigation, a new circRNA named circRNA-0100 was identified from the SHF of cashmere goats, but its function is unknown. In this work, we found that circRNA-0100 exhibited significantly higher expression at anagen SHF bulge than its counterpart at telogen in cashmere goats. Based on the use of both overexpression and siRNA interference assays, our data indicated that circRNA-0100 promoted the differentiation of cashmere goat SHF stem cells (SHF-SCs) into hair follicle lineage, which was evaluated by analyzing the transcriptional level changes of six indicator genes in SHF-SCs of cashmere goats. Using the RNA pull-down technique, we showed that circRNA-0100 served as "molecular sponges" of miR-153-3p in SHF-SCs. Through the use of dual-luciferase reporter assays, our data indicated that circRNA-0100 positively regulated the transcriptional expression of the KLF5 gene via the miR-153-3p-mediated pathway. Ultimately, we showed that circRNA-0100 promoted the differentiation of SHF-SCs into hair lineage, which might be achieved via sequestering miR-153-3p to heighten the KLF5 expression in SHF-SCs of cashmere goats. Our results provide novel scientific evidence for revealing the potential molecular regulatory mechanisms on the differentiation of SHF-SCs into hair lineage in cashmere goats.
环状RNA(circRNAs)具有稳定的结构,是一种没有5'和3'游离末端的共价闭合环。它们可以作为“miRNA海绵”来调节其靶基因的表达。人们认为circRNAs参与了绒山羊次级毛囊(SHF)的发育。在我们之前的研究中,从绒山羊的SHF中鉴定出一种名为circRNA - 0100的新circRNA,但其功能尚不清楚。在这项工作中,我们发现circRNA - 0100在绒山羊生长期SHF隆突处的表达明显高于其在休止期的对应部位。基于过表达和siRNA干扰实验,我们的数据表明circRNA - 0100促进了绒山羊SHF干细胞(SHF - SCs)向毛囊谱系的分化,这是通过分析绒山羊SHF - SCs中六个指示基因的转录水平变化来评估的。使用RNA下拉技术,我们表明circRNA - 0100在SHF - SCs中作为miR - 153 - 3p的“分子海绵”。通过双荧光素酶报告基因实验,我们的数据表明circRNA - 0100通过miR - 153 - 3p介导的途径正向调节KLF5基因的转录表达。最终,我们表明circRNA - 0100促进了SHF - SCs向毛发谱系的分化,这可能是通过隔离miR - 153 - 3p以提高绒山羊SHF - SCs中KLF5的表达来实现的。我们的结果为揭示绒山羊SHF - SCs向毛发谱系分化的潜在分子调控机制提供了新的科学证据。