RNA sequencing of adult cardiomyocyte cyclin C deletion

The goal of this study was to identify differentially expressed genes in hearts with cardiomyoycte deletion of cyclin c compared to cyclin c floxed controls. RNA from three, six week old male hearts from cyclin C fl/fl and cyclin C fl/fl; aMHC cre was isolated. The University of Iowa Institute of Human Genetics, Genomics Division, generated polyA-enriched stranded RNA libraries followed by RNAseq using the Illumina HiSeq platform. Raw sequence reads were analyzed with BaseSpace (www.illumina.com) by aligning reads to the Mus musculus mm10 genome using the TopHat Alignment app. Transcripts were assembled and significant differentially expressed genes were determined with the Cufflinks Assembly and DE app using a false discovery rate <0.05.本研究旨在对比周期蛋白C（cyclin C） flox对照小鼠心脏，鉴定周期蛋白C心肌细胞敲除小鼠心脏中的差异表达基因。研究提取了6周龄雄性cyclin C fl/fl及cyclin C fl/fl; aMHC cre小鼠的心脏总RNA，每组各3份样本。由爱荷华大学人类遗传学研究所基因组学部构建polyA富集链特异性RNA文库，并采用Illumina HiSeq测序平台完成RNA测序。原始测序读段通过BaseSpace平台（www.illumina.com）进行分析：借助TopHat比对工具将读段比对至小家鼠（Mus musculus）mm10参考基因组；随后通过Cufflinks转录本组装与差异表达分析工具，以错误发现率（false discovery rate, FDR）<0.05为阈值，完成转录本组装及显著差异表达基因的筛选。