A versatile and efficient method for detecting tRNA-derived fragments

IF 2.3 3区 生物学 Q3 BIOCHEMICAL RESEARCH METHODS
Mei Yang , Yongzhen Mo , Daixi Ren , Yan Hu , Yiting Tian , Zhaoyang Zeng , Wei Xiong
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引用次数: 0

Abstract

Recently, it has been discovered surprisingly that tRNA can be cleaved into specific small fragments under certain conditions. Most importantly, these tRNA-derived fragments (tRFs) participate in the regulation of gene expression, playing pivotal roles in various physiological and pathological processes and thus attracting widespread attention. Detecting tRF expression in tissues and cells often involves using tRF-specific stem-loop primers for reverse transcription. However, the high specificity offered by this method limits it to transcribing only one specific tRF sequence per reaction, necessitating separate reverse transcription and qPCR steps for multiple tRFs, leading to substantially increased time and resource consumption. This becomes especially challenging in precious samples with limited RNA availability. To address these issues, there is an urgent need for a universal and cost-effective tRF identification method. This study introduces a versatile tRF detection approach based on the uniform polyadenylation of all tRFs, allowing reverse transcription with a universal oligo(dT) primer. This method enables simultaneous reverse transcription of all target tRFs in one reaction, greatly facilitating subsequent qPCR analysis. Furthermore, it demonstrates exceptional sensitivity and specificity, offering significant value in tRF-related research.

检测 tRNA 衍生片段的多功能高效方法。
最近,人们惊奇地发现,tRNA 在某些条件下可以被裂解成特定的小片段。最重要的是,这些 tRNA 衍生的片段(tRFs)参与了基因表达的调控,在各种生理和病理过程中发挥着关键作用,因此受到广泛关注。检测组织和细胞中 tRF 的表达通常需要使用 tRF 特异性茎环引物进行反转录。然而,这种方法的高特异性限制了它在每个反应中只能转录一个特异的 tRF 序列,因此必须对多个 tRF 进行单独的反转录和 qPCR 步骤,从而大大增加了时间和资源消耗。这对核糖核酸供应有限的珍贵样本尤其具有挑战性。为解决这些问题,迫切需要一种通用且经济有效的 tRF 鉴定方法。本研究介绍了一种多功能 tRF 检测方法,该方法基于所有 tRF 的统一多腺苷酸化,允许使用通用的寡聚(dT)引物进行反转录。这种方法能在一个反应中同时反转录所有目标 tRFs,大大方便了后续的 qPCR 分析。此外,它还具有极高的灵敏度和特异性,为 tRF 相关研究提供了重要价值。
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来源期刊
Molecular and Cellular Probes
Molecular and Cellular Probes 生物-生化研究方法
CiteScore
6.80
自引率
0.00%
发文量
52
审稿时长
16 days
期刊介绍: MCP - Advancing biology through–omics and bioinformatic technologies wants to capture outcomes from the current revolution in molecular technologies and sciences. The journal has broadened its scope and embraces any high quality research papers, reviews and opinions in areas including, but not limited to, molecular biology, cell biology, biochemistry, immunology, physiology, epidemiology, ecology, virology, microbiology, parasitology, genetics, evolutionary biology, genomics (including metagenomics), bioinformatics, proteomics, metabolomics, glycomics, and lipidomics. Submissions with a technology-driven focus on understanding normal biological or disease processes as well as conceptual advances and paradigm shifts are particularly encouraged. The Editors welcome fundamental or applied research areas; pre-submission enquiries about advanced draft manuscripts are welcomed. Top quality research and manuscripts will be fast-tracked.
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