小柯机器人

西班牙巴塞罗那科技学院Eva Maria Novoa团队近期取得重要工作进展，他们通过开发出纳米孔RNA测序方法，对tRNA丰度和修饰进行定量分析。相关研究成果2023年4月6日在线发表于《自然—生物技术》杂志上。

据介绍，转运核糖核酸（tRNA）在蛋白质翻译中起着核心作用。研究它们一直很困难，部分原因是缺乏一种同时量化其丰度和化学修饰的简单方法。

研究人员介绍了Nano-tRNAseq，这是一种基于纳米孔的方法，用于对天然tRNA群体进行测序，在单个实验中提供tRNA丰度和修饰动力学的定量估计。研究人员发现，默认的纳米孔测序设置会丢弃绝大多数tRNA读数，导致测序产量低，并且基于转录物长度对tRNA丰度的表示有偏差。对原始纳米孔电流强度信号的再处理导致恢复的tRNA读数的数量增加12倍，并能够重新获得准确的tRNA丰度。随后，研究人员将Nano-tRNAseq应用于酿酒酵母tRNA群体，揭示了同一分子内不同tRNA修饰类型之间的交流和相互依赖性，以及tRNA群体对氧化应激的反应变化。

附：英文原文

Title: Quantitative analysis of tRNA abundance and modifications by nanopore RNA sequencing

Author: Lucas, Morghan C., Pryszcz, Leszek P., Medina, Rebeca, Milenkovic, Ivan, Camacho, Noelia, Marchand, Virginie, Motorin, Yuri, Ribas de Pouplana, Llus, Novoa, Eva Maria

Issue&Volume: 2023-04-06

Abstract: Transfer RNAs (tRNAs) play a central role in protein translation. Studying them has been difficult in part because a simple method to simultaneously quantify their abundance and chemical modifications is lacking. Here we introduce Nano-tRNAseq, a nanopore-based approach to sequence native tRNA populations that provides quantitative estimates of both tRNA abundances and modification dynamics in a single experiment. We show that default nanopore sequencing settings discard the vast majority of tRNA reads, leading to poor sequencing yields and biased representations of tRNA abundances based on their transcript length. Re-processing of raw nanopore current intensity signals leads to a 12-fold increase in the number of recovered tRNA reads and enables recapitulation of accurate tRNA abundances. We then apply Nano-tRNAseq to Saccharomyces cerevisiae tRNA populations, revealing crosstalks and interdependencies between different tRNA modification types within the same molecule and changes in tRNA populations in response to oxidative stress.

DOI: 10.1038/s41587-023-01743-6

Source: https://www.nature.com/articles/s41587-023-01743-6

Nature Biotechnology：《自然—生物技术》，创刊于1996年。隶属于施普林格·自然出版集团，最新IF：68.164
官方网址：https://www.nature.com/nbt/
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