High-resolution quantitative profiling of tRNA abundance and modification status in eukaryotes by mim-tRNAseq
Measurements of cellular tRNA abundance are hampered by pervasive blocks to cDNA synthesis at modified nucleosides and the extensive similarity among tRNA genes. We overcome these limitations with modification-induced misincorporation tRNA sequencing (mim-tRNAseq), which combines a workflow for full-length cDNA library construction from mature tRNA with a simple-to-use computational analysis toolkit. Our method accurately captures tRNA abundance and modification status in multiple eukaryotes and is applicable to any organism with a known genome. Overall design: Development of method for high-throughput sequencing, quantitation and data analysis of tRNA pools in multiple eukaryotic species. Evidence of application for assesing differential expression of tRNAs between two human cell lines, investigating tRNA charging fraction, and extensive modification analysis.