# CRG's tRIBO-seq nanopore method reads tRNAs on active ribosomes and shows how stress changes them

- Published: 2026-11-06 03:39 UTC
- Section: Science › Instruments & experiments
- Tags: tRIBO-seq, ribosome-associated tRNA, nanopore tRNA sequencing, tRNA modifications, tRNA fragmentation, translation regulation, leucine deprivation, methionine starvation, arsenite stress, Centre for Genomic Regulation
- URL: https://realtech.media/news/rt-12b4b5e487ebeeda3ad34b30_crg_s_tribo/

**Barcelona's CRG and IMMAGINA Biotechnology developed tRIBO-seq, which sequences tRNAs from translating ribosomes; under stress these differ from the cell's total tRNA pool.**

![CRG's tRIBO-seq nanopore method reads tRNAs on active ribosomes and shows how stress changes them](https://sgzwpqqelzmajkcubgkl.supabase.co/storage/v1/object/public/rt-media/2026/11/b810d8fe7da76302.jpg)
*NASA, Wikimedia Commons, Public domain*

Researchers at the Centre for Genomic Regulation (CRG) in Barcelona, together with the Italian company IMMAGINA Biotechnology, developed tRIBO-seq, a nanopore-based method that selectively captures transfer RNAs (tRNAs) from actively translating ribosomes, according to a paper in Nature Methods [1]. From a single experiment the method returns tRNA abundance, chemical modifications and fragmentation [1]. The study was led by ICREA Research Professor Eva Novoa; the first author is Mie Monti.

## Why ribosome-associated tRNAs matter

During translation, tRNAs deliver specific amino acids to the ribosome in line with the sequence encoded by the messenger RNA [1]. Their contribution to the modulation of translation remains poorly understood, mainly because there were no methods to characterize tRNA abundance and modifications from actively translating ribosomes [1].

Until now, tRNAs were usually studied by measuring all of them in a cell at once, on the assumption that this reflects the molecules doing the work. Monti said the new results show this is not necessarily the case, particularly under stress.

The method fishes out ribosomes that are busy making proteins and then reads the tRNAs inside one by one by nanopore sequencing. According to the CRG, it needs a single dish of cells and about a day and a half, while conventional methods require about a week and specialist equipment.

## What changes under stress

Using tRIBO-seq, the authors found that ribosome-associated tRNAs (ribo-tRNAs), but often not total tRNAs, are significantly altered under stress [1]. The changes depended strongly on the type of stress [1].

Viral infection and leucine deprivation changed ribo-tRNA abundances [1]. In leucine-starved cells the team saw tRNAs accumulating on the ribosome, although a loss of tRNAs had been expected, Monti said. Methionine starvation caused a dramatic loss of methyl-based tRNA modifications [1].

Arsenite exposure altered neither tRNA abundances nor modification patterns. Instead, it caused major fragmentation in selected subsets of ribo-tRNAs [1].

The authors describe tRIBO-seq as a robust and reproducible approach for mapping the tRNA landscape, covering abundance, modification and fragmentation in both total and actively translating tRNA populations [1].

## Sources

1. [Nature Methods](https://doi.org/10.1038/s41592-026-03277-6)
2. [news-medical.net](https://www.news-medical.net/news/20261105/Novel-method-identifies-active-tRNAs-during-cellular-protein-synthesis.aspx)

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Source: RealTech, realtech.media — https://realtech.media/news/rt-12b4b5e487ebeeda3ad34b30_crg_s_tribo/