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Updated: Sep 16, 2025

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纳米3P-seq:在单分子分辨率下绘制编码和非编码的转录组图

Oguzhan Begik¹, Leszek P Pryszcz¹, Adnan Muhammad Niazi²

¹Centre for Genomic Regulation (CRG), The Barcelona Institute of Science and Technology, Barcelona, Spain.

Nature protocols

|July 8, 2025

概括

这项研究介绍了纳米孔3'终端捕获测序 (Nano3P-seq) 用于分析RNA多A尾部动态. 该方法准确量化了RNA丰富度,尾巴长度和各种RNA类型的组成.

科学领域:

分子生物学分子生物学
基因组学就是基因组学.
生物信息学是一种生物信息学.

背景情况:

RNA多化对RNA处理至关重要,影响稳定性,翻译和衰变.
多甲尾长是mRNA功能和调节的一个关键决定因素.
目前用于分析多A尾的方法在范围或分辨率上可能受到限制.

研究的目的:

介绍纳米孔3'终端捕获测序 (Nano3P-seq) 的详细协议.
引入PolyTailor软件,用于分析多的尾巴长度和组成.
为了使单分子分辨率分析的RNA尾巴动态跨各种RNA类型.

主要方法:

使用纳米孔测序开发和验证Nano3P-seq协议.
使用基于模板切换的方法从3'端对cDNA进行测序.
使用PolyTailor软件进行数据分析和预测尾巴特征.

主要成果:

纳米3P-seq准确地估计了转录的丰度,多个A尾的长度和组成.
该方法成功分析了编码和非编码RNA,包括mRNA,snoRNA和rRNA.
该协议与R10.4流细胞兼容,可以在一天内完成.

结论:

纳米3P-seq为研究RNA尾部动态提供了一个全面和可重复的方法.
该协议可适应各种RNA样品准备方法.
这种技术增强了对转录组范围内的尾巴异质性的研究.

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相关概念视频

RNA-seq

RNA-seq

RNA sequencing, or RNA-Seq, is a high-throughput sequencing technology used to study the transcriptome of a cell. Transcriptomics helps to interpret the functional elements of a genome and identify the molecular constituents of an organism. Additionally, it also helps in understanding the development of an organism and the occurrence of diseases.
Before the discovery of RNA-seq, microarray-based methods and Sanger sequencing were used for transcriptome analysis. However, while...