Next Generation Sequencing Facilitates Quantitative Analysis of RBM22-silencing and Control Primary Neonatal Mouse Cardiomyocytes Transcriptomes

Purpose:The purpose of this study is to identify genes that are either activated or silenced in cardiomyocytes treated with siRNA targeting RBM22 (si-Rbm22) compared to control siRNA (si-Control). Gene expression differences between the two samples were identified using transcriptome profiling (RNA-seq) analysis. Methods: Primary neonatal mouse cardiomyocytes were isolated from neonatal C57BL/6J mice at postnatal day 1.Cardiomyocytes were stained to confirm the expression of cTnT. Cells with purity >97.5% were used for subsequent experiments. Neonatal cardiomyocytes in primary culture were transfected with either siRNA targeting Rbm22 (si-Rbm22) or a non-targeting control siRNA (si-Control) for 48 h, of which RNA profiles were generated by deep sequencing using Illumina. Results: We mapped about 10 million sequence reads per sample to the mouse genome and identified numerous genes with significant mRNA variation between cardiomyocytes transfected with the indicated siRNAs. Overall design: mRNA profiles of neonatal mouse cardiomyocytes transfected with control siRNA or Rbm22 siRNA were generated by deep sequencing.