Small RNA Deep Sequencing Analysis of Mouse T Helper Cell (Th) Subsets

Small non-coding RNAs, esp. microRNAs (miRNAs), play critical roles in a large diversity of biological processes. The goal of this study is to perform small RNA deep sequencing analysis of mouse Th subsets and to identify miRNAs that are highy and specifically expressed in each subset. Overall design: Naive T cells from several transcriptional reporter mice (Ifngyfp, Il17frfp and Foxp3gfp) were differentiated into appropriate Th subsets in vitro and harvested on day 4-5. Cell sorting was performed using a FACSAria cell sorter to obtain reporter gene-positive cells. Total RNA was extracted using the TRIzol reagent and subjected to standard small RNA deep sequencing analysis.