Table_2_Transcriptomic Analysis Reveals the Involvement of lncRNA–miRNA–mRNA Networks in Hair Follicle Induction in Aohan Fine Wool Sheep Skin.XLSX

Long non-coding RNAs (lncRNA) and microRNAs (miRNA) are new found classes of non-coding RNAs (ncRNAs) that are not translated into proteins but regulate various cellular and biological processes. In this study, we conducted a transcriptomic analysis of ncRNA and mRNA expression in Aohan fine wool sheep (AFWS) at different growth stages (embryonic day 90, embryonic day 120, and the day of birth), and explored their relationship with wool follicle growth. In total, 461 lncRNAs, 106 miRNAs, and 1,009 mRNAs were found to be differentially expressed during the three stages of wool follicle development. Gene Ontology (GO) enrichment and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analysis were performed to clarify the roles of the differentially expressed lncRNA, miRNA, and mRNA in the different stages of wool follicle development. Quantitative real-time PCR (qRT-PCR) was used to validate the results of RNA-seq analysis. lncRNA (MSTRG.223165) was found to act as a competing endogenous RNA (ceRNA) and may participate in wool follicle development by acting as an miR-21 sponge. Network prediction implicated the MSTRG.223165-miR-21-SOX6 axis in the wool follicle development. The targeting relationships of miR-21 with SOX6 and MSTRG.223165 were validated in dual-luciferase assays. This is the first report indicating the association of the lncRNA–miRNA–mRNA network with wool follicle development in AFWS. This study provides new insights into the regulation of the wool follicle growth and represents a solid foundation for wool sheep breeding programs.

长链非编码RNA（long non-coding RNAs，lncRNA）与微小RNA（microRNAs，miRNA）是新近发现的非编码RNA（non-coding RNAs，ncRNA）类别，此类RNA不翻译成蛋白质，却可调控多种细胞及生物学过程。本研究对不同生长阶段（胚胎第90天、胚胎第120天及出生当天）的敖汉细毛羊（Aohan fine wool sheep，AFWS）开展非编码RNA及mRNA表达的转录组分析，探究其与毛囊生长的关联。结果显示，在毛囊发育的三个阶段中，共鉴定得到461个差异表达的lncRNA、106个差异表达的miRNA以及1009个差异表达的mRNA。通过基因本体（Gene Ontology，GO）富集分析与京都基因与基因组百科全书（Kyoto Encyclopedia of Genes and Genomes，KEGG）通路分析，明确了差异表达的lncRNA、miRNA及mRNA在毛囊发育不同阶段的功能。采用实时荧光定量PCR（quantitative real-time PCR，qRT-PCR）对RNA测序（RNA-seq）的分析结果进行验证。研究发现，长链非编码RNA MSTRG.223165可作为内源竞争RNA（competing endogenous RNA，ceRNA），通过充当miR-21的分子海绵参与毛囊发育。网络预测结果显示，MSTRG.223165-miR-21-SOX6调控轴参与毛囊发育过程。双荧光素酶报告实验验证了miR-21与SOX6、MSTRG.223165的靶向结合关系。本研究首次揭示了敖汉细毛羊中lncRNA–miRNA–mRNA调控网络与毛囊发育的关联，为毛囊生长调控机制提供了全新视角，同时为毛用绵羊育种工作奠定了坚实的理论基础。