绒山羊胎儿期毛囊发生发育关键circRNA的鉴定及功能预测分析

Identification and functional prediction analysis of key circRNA for hair follicle development in the fetal period of cashmere goat

为探究调控绒山羊毛囊发生发育阶段关键circRNA及其功能,基于课题组前期得到的内蒙古绒山羊胎儿期(45、55、65和75d)皮肤组织circRNA表达谱以及各比对组中差异表达circRNA的基础,根据不同时期毛囊形态发生发育情况筛选了毛囊发生发育相关的circRNA,利用生物信息学分析毛囊发生发育关键circRNA,并对其进行qRT-PCR试验;运用TargetScan和miRanda预测关键circRNA靶向的miRNA以及靶向miRNA的靶基因,分析了22个内含子circRNA宿主基因的富集情况,探究circRNA的功能作用。结果表明:1)通过生物信息学分析,circRNA2049、circRNA5681、circRNA2225和circRNA3411均由外显子组成,为外显子circRNA。2)qRT-PCR显示,circRNA2049、circRNA2225、circRNA3411和circRNA5681在绒山羊胎儿期4个时期(45、55、65和75d)中均显著差异表达(P<0.05)。3)功能分析显示,外显子circRNA可能竞争性结合miRNA,以circRNA-miRNA-mRNA调控网络来调控毛囊的发生发育,同时内含子circRNA宿主基因也可能影响毛囊的发生发育,具体作用机制需后续探究。综上,本研究筛选鉴定了绒山羊毛囊发生发育关键circNRA,并初步探究了其功能,为解析circRNA在绒山羊毛囊发育过程中的分子机制提供了理论基础,为研究绒毛生长发育提供了新的方向。

In order to explore the key circRNAs and their functions at the hair follicle development stage of Inner Mongolia Cashmere goat during fetal period,this study is based on the circRNA expression profile of skin tissue of Inner Mongolia Cashmere goat during fetal period(45,55,65 and 75 d)obtained by the research group earlier and the basis of differential expression of circRNA in each comparison group.According to the hair follicle morphogenesis and development in different periods,the circRNAs related to hair follicle genesis and development were screened,the key circRNAs of hair follicle genesis and development were analyzed by bioinformatics,and the qRT-PCR experiment was carried out.Targetscan and miRanda were adopted to predict the miRNAs targeted by key circRNAs and the target genes targeting miRNAs.The enrichment of 22 intron circRNA host genes was analyzed to explore the functional role of circRNAs.The results showed that:1)Bioinformatics analysis discovered that circRNA2049,circRNA5681,circRNA225 and circRNA3411 were all composed of exons,which were exon circRNA.2)The results of qRT-PCR showed that circRNA2049,circRNA2225,circRNA3411 and circRNA5681 were differentially expressed in the four fetal stages of cashmere goats(45,55,65 and 75 d)(P<0.05).3)In terms of functional analysis,it was found that exon circRNA might competitively bind miRNA to regulate the morphogenesis and development of hair follicles with circRNAmiRNA-mRNA regulatory network.At the same time,its host gene may also affect the occurrence and development of hair follicles.The specific mechanism still needs to be explored in subsequent experiments.In conclusion,this study screened and identified the key circRNA in the morphogenesis and development of cashmere goat hair follicle and preliminarily explored its function,which provides a theoretical basis for analyzing the molecular mechanism of circRNA in the development of cashmere goat hair follicle,and provides a new direction for the study of cashmere growth and development.