Characterization Of Lncrna And Function Analysis Inmyoblast Of Meat Sheep

Sheep is an important economic animals and production trait has become the main direction of sheep breeding. There are significant differences between different sheep breeds on development mechanism which can help us to discover unknown genes. Lnc RNAs are a class of non-coding RNAs that are longer than 200 nt and have an important role in mammal development which have a characterization of temporal differential expression.We chose two different sheep varieties which had significant differences in the development, Ujimqin and Texel, to analyze the gastrocnemius using the next generation sequencing technologies at 85 d and 120 d of gestation, newborn and one-year old, respectively, and to find the internal mechanism of productivity difference. We studied lnc RNA which was screened out, TCONS₀0102859, by sheep myoblast. The main findings are followings: 1. Lnc RNA sequencing and descriptive analysisWe filtered average of 2.25 Gb of data per sample, including a minimum amount of data at 85 d fetus, and the quality of data was reliable. We find more than 89% of over all mapping rate on genome and obtain replies genomic data filtering, to obtained 2002 candidates lnc RNAs. There was no chromosome preference and a large length across, but 78.5% of lnc RNA has a length of < 2000 nt. Lnc RNA has 2.8 exons average and 90% lnc RNA has fewer than 4 exons. We found 1967 new transcripts. 2. Annotation of target genes of lnc RNAWe found that lnc RNA may be associated with binding and catalysis, metabolic processes, pigmentation and reproductive performance, and so on through GO analysis of target genes. We found large number of genes associated with signal transduction, infectious disease, cancer, transcription and translation through KEGG analysis. 3. Differential expression analysis of lnc RNA at each developmental stage between Ujimqin and TexelLnc RNAs analysis of differential expression at each developmental stage between these two breeds, we found that enrichment results were significant difference. We enriched MAPK signaling pathway and Wnt signaling pathway which were associated with muscle development, tumorigenesis and so on at 85 d. Cardiac-related pathways were enriched at 120 d. Phosphatidylinosital signaling system and calcium signaling pathway were enriched at nascent stage and we found nicotinate and nicotinamide metabolism at one-year-old sheep. Ten lnc RNAs were found different expression at all developmental stages of the two species. There are three lnc RNAs, TCONS₀0051148, TCONS₀0079077 and TCONS₀0078553, were corresponding to the internal genes which were ANKRD10, FGF2 and LOC101105780, respectively. We found TCONS₀0102859 expressed in Texel only and gradually increased as time goes on. 4. Preliminary analysis of TCONS₀0102859 on sheep myoblastLentivirus vector that overexpress TCONS₀0102859 was constructed and applied on sheep myoblast. High transfection efficiency and overexpression efficiency were found. Myo D, Myo G and Mef2 c were upregulated and MSTN were downregulated in transfected myoblast. That was the preliminary evidence that TCONS₀0102859 may have a role both on proliferation and differentiation of myoblast.The identification and function analysis in myoblast of sheep lnc RNAs will further promote the study of lnc RNA functions and gene regulatory mechanisms.