Male and female mouse heart miRNA sequencing

Purpose: The goal of this study is to identify the sex differential miRNA tanscriptome between male and female adult (4-6 wk. old) mouse hearts using small RNA-seq. Methods: miRNA profiles of male and female mouse hearts were generated by sequencing, n=4 for each sex, using Illumina HiSeq2500. The sequence reads were aligned to the mm10 reference genome using STAR via the bcbio-nextgen RNA-sequencing pipeline. Differential gene expression was determined by genwiz. Results: 11 miRNAs were differentially expressed between male and female mice [ P value <0.05, |log2(Fold Change)| > 0.5], with 9 more highly expressed in female hearts and 2 more highly expressed in male hearts. Overall design: Male and female adult hearts (4-6 wks), n=4 for each sex, were dissected in cold PBS and immediately homogenized in Trizol solution, and were kept in -20C until use. Small RNA from individual hearts was extracted using the miRVANA miRNA isolation kit (Thermo-Fisher AM1561). The concentration and quality of purified RNA were assessed by Tape Station (Agilent). Isolated small RNA samples were used for library prep and sequencing regardless of RIN score because small RNA bands were expected. The samples were enriched in RNAs <200nt.