RNA-seq of metabolically labelled RNA by 4-thiouracil from mouse tissues

To identify cell type-specific RNA without cell sorting, mice expressing UPRT in a specific cell type were exposed to 4-thiouracil to label RNA only in the UPRT-expressing cells. Total RNA was extracted from tissues and treated with iodoacetamide followed by RNA-seq library preparation. The reads generated from 4-thiouracil-containing transcripts should contain T to C mismatches as a result of 4-thiouracil incorporations and thus can be identified as the transcripts generated in the UPRT-expressing cells.