Identification And Functional Of Differentially Expressed LncRNA Target Genes During Secondary Follicle Initiation In Fine-wool Sheep

Secondary hair follicle(SF) initiation of sheep is depend on a series of complex inter-cellular signaling pathway interaction between epiderms and dermis. Signaling pathway in any one ligand, receptor genetic mutations, epigenetic modification and post-translational modification of the protein changes will affect sheep SF occurs.Thus, in addition to the current level of interest in the protein-coding m RNA genes,non-coding of long-chain RNA(long noncoding RNA, lnc RNA) as a new type of gene expression regulatory factors, it’s also to be focused. A lot of research showed that Lnc RNA plays an important role in organofaction, cell differentiation and disease occurrence, but whether Lnc RNA influence the hair follicle morphogenesis is very less. Therefore, in the early we use RNA-seq sequencing technology for Alpine Merino sheeep hair foliicle initiation of skin tissue were sequenced, this study depend on this basis to screen differences Lnc RNA. And through bioinformatics and luciferase reporter gene system to prediction and verification Lnc RNA target gene to discuss Lnc RNA mechanism in fine wool hair follicle morphogenesis.(1) Used CNCI, Phylo CSF and pfam protein domain to analyze the sequencing data of fine wool sheep skin during the initiation, It found that 884 new Lnc RNAs are relate to follicle morphogenesis. Compare with by Edge R, 15 Lnc RNA prominent different expression gene were found. We predicted these 15 Lnc RNA,13 Lnc RNA exist potential cis or trans target gene. And through the GO and KEGG enrichment analysis, the 13 Lnc RNA target gene significant enrichment to NOD-like receptor,Leishmaniasis and NF-kappa B sighaling pathways. Explained that these difference expression of Lnc RNA may be involved in the fine-wool sheep follicle morphogenesis through signal pathway.(2) Use BLAST compare the Lnc RNA005698, Lnc RNA000629 with mature mi RNA in sheep mi RNA Base Database, It found that Lnc RNA005698 and oar-mi R-3955-5p has high coherence seed region, and them binding minimum free energy(MFE=34.4 kcal/mol) at a low level. Indicated Lnc RNA005698 may beoar-mi R-3955-5p competing endogenous RNA(ce RNA). The relationship between Lnc RNA005698 and oar-mi RNA-3955-5p was validated by the luciferase reporter gene system, which indicated that between oar-mi RNA-3955-5p and Lnc RNA005698 existence the relationship of target. Use the RT-PCR to analyze the expression profile of Lnc RNA005698 and oar-mi RNA-3955-5p in sheep heart, liver, spleen, lung,kidney tissues. The result showed that Lnc RNA005698 and oar-mi RNA-3955-5p is also has the relationship of target。To sum up, Lnc RNA005698 may plays an important role in the process of fine-wool sheep hair follicle, and may by adjusting the oar-mi RNA-3955-5p to regulate the fine-wool sheep hair follicle morphogenesis. The study on the regulation of wool bursa morphogenetic Lnc RNA for inquiry of non coding RNA in sheep hair follicle morphogenesis occurs mechanism and provide a theoretical reference.