RNA-seq muscle (AR)

Identify the genes modulated by Androgen Receptor. We selectively knocked-out AR in myofibers by crossing AR floxed mice to HSA-Cre mice. RNA from gastrocnemius muscle of 9 week-old control (AR floxed mice) and knock-out mice was extracted with trizol according to the manufacturer protocol. RNA samples were sequenced by the standard Illumina protocol to create raw sequence files (.fastq files). We annotated these reads to the mm10 build of the mouse genome using HOMER. The aligned reads were counted with the homer software (analyze RNA) and DEG's were identified using EdgeR and DESeq.本研究旨在鉴定受雄激素受体（Androgen Receptor, AR）调控的基因。通过将AR floxed小鼠与HSA-Cre工具小鼠交配，实现肌纤维中AR的特异性敲除。按照试剂厂商的操作规程，使用TRIzol试剂提取9周龄对照组（AR floxed小鼠）与AR敲除小鼠的腓肠肌总RNA。采用标准Illumina测序流程对RNA样本进行测序，生成原始测序数据文件（.fastq格式）。利用HOMER工具将测序reads比对注释至小鼠基因组mm10组装版本，随后通过HOMER软件的analyze RNA模块对比对reads进行计数，并借助EdgeR与DESeq软件鉴定差异表达基因（Differentially Expressed Genes, DEGs）。