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 endogenously modified tRNA with a comprehensive and user-friendly computational analysis toolkit. Our method accurately captures tRNA abundance and modification status in yeast, fly, and human cells and is applicable to any organism with a known genome. We applied mim-tRNAseq to discover a dramatic heterogeneity of tRNA isodecoder pools among diverse human cell lines and a surprising interdependence of modifications at distinct sites within the same tRNA transcript.

Automated summary

The study overcame the challenges in measuring cellular tRNA abundance by using modification-induced misincorporation tRNA sequencing (mim-tRNAseq) method, accurately capturing tRNA abundance and modification status in cells. The research revealed significant heterogeneity of tRNA isodecoder pools among diverse human cell lines and surprising interdependence of modifications at distinct sites within the same tRNA transcript.