RNA_Sequencing Facilitates Quantitative Analysis of macrophages transcriptomes from Wild Type and miR_7 deficiency mice Transcriptomes

Purpose: The goals of this study are to analyze the differential gene expression of miR_7 deficiency mice macrophages before and after activation, and screen out miR_7 candidate target genes. Total RNA of macrophages (from WT or miR_7 deficiency mice) stimulated with 100 ng/mL LPS for 12 hrs or left unstimulated was purified using TRIzol reagent. Total RNA of each sample was submitted to the TSRI Next Generation Sequencing Core for deep sequencing. Total RNA was enriched for polyA_containing RNAs and deep sequencing of those samples was performed on a HiSeq 2000 (Illumina). Data analysis was performed with Pipeline Software (Casava v1.8.2) and Cutadapt. Alignment to the mouse genome was performed with mRNA_Seq TopHat v2.0.13 with Bowtie2, and the exon annotation retrieved with Partek. Total read counts for each gene was calculated and used to identify protein coding genes downregulated by miR_7.