The abundance and modification profiles of tRNAs are dynamic and contribute to their biological function. The quantification of tRNA abundance and chemical modifications has been challenging using currently available approaches. Lucas et al. developed a nanopore-based, high-throughput sequencing method, named Nano-tRNAseq, that enables accurate measurement of both the abundance and modification dynamics of tRNAs simultaneously at single-molecule resolution. The authors modified the direct RNA sequencing protocols developed by Oxford Nanopore Technologies with optimized mapping settings and custom MinKNOW parameters, which led to a 12-fold increase in the number of sequenced and mapped tRNA reads, compared with the sequencing yield using default settings. Nano-tRNAseq can quantify tRNA abundance and capture modification interdependencies. Applying Nano-tRNAseq to Saccharomyces cerevisiae tRNA populations subjected to environmental exposures revealed systematic de-adenylation of the 3′ CCA tail of tRNA molecules during oxidative stress. Nano-tRNAseq might be applied to various conditions and advance our understanding of the small RNA transcriptome and native RNA modifications as well as their function in human diseases.

tRNA 的丰度和修饰谱是动态的，有助于其生物学功能。使用当前可用的方法，tRNA 丰度和化学修饰的量化一直具有挑战性。卢卡斯等人。开发了一种基于纳米孔的高通量测序方法，名为 Nano-tRNAseq，能够以单分子分辨率同时准确测量 tRNA 的丰度和修饰动力学。作者使用优化的映射设置和自定义 MinKNOW 参数修改了 Oxford Nanopore Technologies 开发的直接 RNA 测序方案，与使用默认设置的测序产量相比，这导致测序和映射的 tRNA 读数数量增加了 12 倍。Nano-tRNAseq 可以量化 tRNA 丰度并捕获修饰的相互依赖性。暴露于环境的酿酒酵母tRNA 种群显示，在氧化应激期间，tRNA 分子的 3' CCA 尾发生了系统性去腺苷酸化。Nano-tRNAseq 可应用于各种条件，并促进我们对小 RNA 转录组和天然 RNA 修饰及其在人类疾病中的功能的理解。