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High-resolution quantitative profiling of tRNA abundance and modification status in eukaryotes by mim-tRNAseq

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Behrens,  Andrew
Nedialkova, Danny / Mechanisms of Protein Biogenesis, Max Planck Institute of Biochemistry, Max Planck Society;

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Rodschinka,  Geraldine
Nedialkova, Danny / Mechanisms of Protein Biogenesis, Max Planck Institute of Biochemistry, Max Planck Society;

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Nedialkova,  Danny D.
Nedialkova, Danny / Mechanisms of Protein Biogenesis, Max Planck Institute of Biochemistry, Max Planck Society;

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Citation

Behrens, A., Rodschinka, G., & Nedialkova, D. D. (2021). High-resolution quantitative profiling of tRNA abundance and modification status in eukaryotes by mim-tRNAseq. Molecular Cell, 81(8), 1802-+. doi:10.1016/j.molcel.2021.01.028.


Cite as: http://hdl.handle.net/21.11116/0000-0008-A38C-2
Abstract
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.