在单细胞中使用STORM-seq有效地对总RNA进行分析
Benjamin K Johnson1, Mary F Majewski1, H Josh Jang1
1Department of Epigenetics, Van Andel Institute, Grand Rapids, MI, USA.
bioRxiv : the preprint server for biology
|June 6, 2025
概括
我们开发了STORM-seq,一种新的单细胞总RNA测序方法. 它克服了当前技术的局限性,使得基因表达的详细分析成为可能,包括非编码RNA和可移植元素.
科学领域:
- 分子生物学分子生物学
- 基因组学就是基因组学.
- 文字转录学 (Transcriptomics) 是一个学科.
背景情况:
- 目前的单细胞RNA测序 (scRNA-seq) 方法在检测非编码转录,实现全长RNA覆盖和解决转录复杂性方面面临挑战.
- 许多scRNA-seq协议需要专门的设备,限制了可访问性和广泛实施.
研究的目的:
- 引入单细胞TOTALRNA-seq小型化 (STORM-seq),这是一个全新的,可访问的协议,用于全面的单细胞总RNA测序.
- 使用标准实验室设备,证明STORM-seq增强转录检测的能力,包括非编码RNA,异形和基因融合.
主要方法:
- STORM-seq采用一种随机六合体原始化,缩的单细胞总RNA测序方法.
- 该协议旨在快速构建图书馆 (一工作日),并且可以作为一个套件进行调整,需要标准实验室设备.
主要成果:
- STORM-seq生成高度复杂的库,使得能够对单细胞中的转录异型和临床相关的基因融合进行可靠的测量.
- 该方法准确地描述了位点级可转移元素 (TE) 和低丰度增强子RNA (eRNA),提供了高分辨率的基因调控网络洞察力.
- 对人类输卵管表皮的应用揭示了由TEs和非编码RNAs影响的原始细胞样群体和中间细胞状态.
结论:
- STORM-seq为详细的单细胞转录组分析提供了一种强大,易于使用的工具,克服了现有的scRNA-seq技术的关键局限性.
- 该协议能够解析复杂的转录组,包括非编码RNA和TE,为剖析单细胞基因调节网络提供了前所未有的分辨率.
相关概念视频
03:21RNA-seq
10.4K
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...
Before the discovery of RNA-seq, microarray-based methods and Sanger sequencing were used for transcriptome analysis. However, while...
10.4K
02:24Ribosome Profiling
3.7K
Ribosome profiling or ribo-sequencing is a deep sequencing technique that produces a snapshot of active translation in a cell. It selectively sequences the mRNAs protected by ribosomes to get an insight into a cell’s translation landscape at any given point in time.
Applications of ribosome profiling
Ribosome profiling has many applications, including in vivo monitoring of translation inside a particular organ or tissue type and quantifying new protein synthesis levels.
The technique...
Applications of ribosome profiling
Ribosome profiling has many applications, including in vivo monitoring of translation inside a particular organ or tissue type and quantifying new protein synthesis levels.
The technique...
3.7K