Transcriptomes of miR-290-295 KO mESCs treated with siPOOLs for three different transcription factors and a negative control. Transcriptomes of miR-290-295 KO mESCs treated with siPOOLs for three different transcription factors and a negative control

In order to identify direct microRNA targets in mouse embryonic stem cells (mESCs), we generated miR-290-295 KO mutant cell lines, assessed their transcriptomes. We identified novel transcription factors (TF) as potential direct and functional targets of miR-290-295. To assess the role of these TFs, we rescued their expression in the miR-290-295 KO cell line using siPOOLs and profiled their transcriptomes. Overall design: Transcriptomesof miR-290-295 KO E14 mESCs after siPOOL treatments for different Transcription Factors were generated in biological duplicates using QuantSeq kit.

为鉴定小鼠胚胎干细胞（mouse embryonic stem cells, mESCs）中microRNA（miRNA）的直接靶标，我们构建了miR-290-295敲除（knockout, KO）突变细胞系，并对其转录组进行了分析。我们鉴定出多个新型转录因子（transcription factor, TF），作为miR-290-295的潜在直接功能靶标。为评估这些转录因子的功能作用，我们利用siPOOLs在miR-290-295 KO细胞系中恢复其表达，并对转录组进行了测序分析。整体实验设计：采用QuantSeq试剂盒，对经不同转录因子siPOOLs处理后的miR-290-295敲除E14小鼠胚胎干细胞的转录组进行了双份生物学重复检测。