Investigation of miRNAs from PBMC in Kangal type Akkaraman sheep with small RNA-seq

Purpose: Aim of this study was to determine the miRNAs associated with average daily gain in Kangal Akkaraman lambs. Depending on the average daily gain, two groups were formed as low average daily gain (LDG) and high average daily gain (HDG). Methods: Peripheral blood mononuclear cells (pbmc) were separated from blood samples obtained from lambs and RNA was isolated with TRI Reagent (Sigma). Isolated total RNAs from LDG (n = 17) and HDG (n = 21) groups were pooled. miRNA libraries (LDG and HDG groups) were prepared with NEBNext® Multiplex Small RNA Library Prep Set for Illumina® (NEB, USA.) and sequenced with Illumina Novaseq. Readings were mapped with the Bowtie program using the Oar_v3.1 sheep reference genome. Differential expression analysis of the two groups was performed using the DESeq R package (1.8.3). Results: Using an optimized data analysis workflow, we mapped about 18 million sequence reads in HDG and 21 million sequence reads n LDG samples to the ovis aries genome (oar_v3.1). In total, 129 known miRNAs were identified, 134 novel miRNAs were predicted, and 40 differentially expressed miRNAs were found between the groups. Conclusions: Our study represents the first detailed analysis of Kangal type Akkaraman sheep PBMC miRNAs with RNA-seq technology. The obtained sequence reads would provide data for futher breeding studies related to average daily gain in Kangal Akkraman sheep. Examination of PBMC miRNA expression in low average daily gain and high average daily gain Kangal Akkaraman lambs.