Profiling of microRNA from skeletal muscle of Bandur sheep using RNA sequencing

Abstract

MicroRNA profiling is a powerful approach for identifying key regulators of molecular functions which control skeletal muscle development, regeneration and function. Information on gene expression and the regulatory factors involved in myogenesis is very limited for Indian sheep. This study reports the identification and characterization of miRNAs from the skeletal muscles of Bandur sheep breed for the first time. Bandur is a consumer favoured, mutton type sheep of India, mainly distributed in Mandya district of Karnataka. Skeletal muscles from four animals of Bandur sheep of similar age, sex and reared under same management conditions were used for RNA sequencing. The total number of reads (15–36 bp) for each library of Bandur sheep ranged from 19,350,000 to 30,000,000. Highly expressed transcripts with an RPKM value of ≥1000 were observed to be 34%, whereas 38% transcripts exhibited RPKM between 100–1000 and 28% had RPKM <100 in Bandur sheep. A total of 110 known mature miRNAs could be identified on comparison with known human and bovine sequences. All the identified miRNAs represented 32 miRNA families and 44 clusters. A total of 499 novel miRNAs were discovered in Bandur sheep. The miRNAs identified in our study were enriched for functions namely cell proliferation, cell differentiation, osteogenesis, lipid metabolism, muscle development, adipocyte differentiation and stress response. Potential gene targets for the identified miRNAs were predicted. Most relevant target genes predicted in our study included MYO5A, SIN3B and NR2F2 which are mainly involved in myogenesis. This study provides information of miRNAs in the skeletal muscle tissue of Bandur sheep.