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...

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Detalles Bibliográficos
Autores principales: Behrens, Andrew, Rodschinka, Geraldine, Nedialkova, Danny D.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Cell Press 2021
Materias:
Technology
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8062790/
https://www.ncbi.nlm.nih.gov/pubmed/33581077
http://dx.doi.org/10.1016/j.molcel.2021.01.028
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author Behrens, Andrew
Rodschinka, Geraldine
Nedialkova, Danny D.
author_facet Behrens, Andrew
Rodschinka, Geraldine
Nedialkova, Danny D.
author_sort Behrens, Andrew
collection PubMed
description 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.
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spelling pubmed-80627902021-04-27 High-resolution quantitative profiling of tRNA abundance and modification status in eukaryotes by mim-tRNAseq Behrens, Andrew Rodschinka, Geraldine Nedialkova, Danny D. Mol Cell Technology 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. Cell Press 2021-04-15 /pmc/articles/PMC8062790/ /pubmed/33581077 http://dx.doi.org/10.1016/j.molcel.2021.01.028 Text en © 2021 The Author(s) https://creativecommons.org/licenses/by-nc-nd/4.0/This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/).
spellingShingle Technology
Behrens, Andrew
Rodschinka, Geraldine
Nedialkova, Danny D.
High-resolution quantitative profiling of tRNA abundance and modification status in eukaryotes by mim-tRNAseq
title High-resolution quantitative profiling of tRNA abundance and modification status in eukaryotes by mim-tRNAseq
title_full High-resolution quantitative profiling of tRNA abundance and modification status in eukaryotes by mim-tRNAseq
title_fullStr High-resolution quantitative profiling of tRNA abundance and modification status in eukaryotes by mim-tRNAseq
title_full_unstemmed High-resolution quantitative profiling of tRNA abundance and modification status in eukaryotes by mim-tRNAseq
title_short High-resolution quantitative profiling of tRNA abundance and modification status in eukaryotes by mim-tRNAseq
title_sort high-resolution quantitative profiling of trna abundance and modification status in eukaryotes by mim-trnaseq
topic Technology
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8062790/
https://www.ncbi.nlm.nih.gov/pubmed/33581077
http://dx.doi.org/10.1016/j.molcel.2021.01.028
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AT nedialkovadannyd highresolutionquantitativeprofilingoftrnaabundanceandmodificationstatusineukaryotesbymimtrnaseq

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