Search research articles

关于 JoVE

概览领导团队博客 JoVE 帮助中心

作者

出版流程编辑委员会范围与政策同行评审常见问题投稿

图书馆员

用户评价订阅访问资源图书馆顾问委员会常见问题

研究

JoVE Journal Methods Collections JoVE Encyclopedia of Experiments 存档

教育

JoVE Core JoVE Business JoVE Science Education JoVE Lab Manual 教师资源中心教师网站

使用条款与条件

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...

您也可能阅读

相关文章

通过共同作者、期刊和引用图与本文相关的文章。

排序

Same author

AURORA A interacts with DICER and SETD2 to promote S-phase progression.

EMBO reports·2026

Same author

Respiratory syncytial viral load drives ciliated cell dedifferentiation and suppresses antiviral immunity.

Science advances·2026

Same author

On the Scope of DCAF1-Recruiting PROTACs Degrading Protein Kinases.

Journal of medicinal chemistry·2026

Same author

Reframing ME/CFS: toward a unified mechanistic model of chronic post-infectious diseases.

Journal of translational medicine·2026

Same author

A murine cytomegalovirus cell cycle regulator (m54.5p) evolved within the conserved viral DNA polymerase gene.

PLoS pathogens·2026

Same author

Minute-scale control of ubiquitin-mediated degradation reveals dynamics of bacterial secreted effector-functions.

Nature communications·2026

Search research articles

相关实验视频

Updated: Jul 2, 2025

Rare Event Detection Using Error-corrected DNA and RNA Sequencing

Rare Event Detection Using Error-corrected DNA and RNA Sequencing

Published on: August 3, 2018

纠正4sU诱导的核酸转换RNA-seq数据中的量化偏差.

Kevin Berg^1,2, Manivel Lodha², Isabel Delazer³

¹Chair of Computational Immunology, University of Regensburg, Regensburg, Germany.

Nucleic acids research

|February 21, 2024

概括

在RNA标记过程中,高度的4-thiouridine (4sU) 可能会影响表达估计. 这项研究开发了计算工具,以纠正核酸转换RNA测序数据中的这些偏差.

更多相关视频

Saccharomyces cerevisiae Metabolic Labeling with 4-thiouracil and the Quantification of Newly Synthesized mRNA As a Proxy for RNA Polymerase II Activity

Saccharomyces cerevisiae Metabolic Labeling with 4-thiouracil and the Quantification of Newly Synthesized mRNA As a Proxy for RNA Polymerase II Activity

Published on: October 22, 2018

Real-time Analysis of Transcription Factor Binding, Transcription, Translation, and Turnover to Display Global Events During Cellular Activation

Real-time Analysis of Transcription Factor Binding, Transcription, Translation, and Turnover to Display Global Events During Cellular Activation

Published on: March 7, 2018

相关实验视频

Last Updated: Jul 2, 2025

Rare Event Detection Using Error-corrected DNA and RNA Sequencing

Rare Event Detection Using Error-corrected DNA and RNA Sequencing

Published on: August 3, 2018

Saccharomyces cerevisiae Metabolic Labeling with 4-thiouracil and the Quantification of Newly Synthesized mRNA As a Proxy for RNA Polymerase II Activity

Saccharomyces cerevisiae Metabolic Labeling with 4-thiouracil and the Quantification of Newly Synthesized mRNA As a Proxy for RNA Polymerase II Activity

Published on: October 22, 2018

Real-time Analysis of Transcription Factor Binding, Transcription, Translation, and Turnover to Display Global Events During Cellular Activation

Real-time Analysis of Transcription Factor Binding, Transcription, Translation, and Turnover to Display Global Events During Cellular Activation

Published on: March 7, 2018

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:

核酸类类似物如4-thiouridine (4sU) 能够对新合成的RNA进行代谢标记.
将4sU转化为C:T不匹配的化学转换允许同时对总和标记的RNA表达进行分析.
潜在的细胞毒性和广泛的4sU标记对RNA-seq表达估计的影响仍未得到充分研究.

研究的目的:

为了研究4 - 氨酸 (4sU) 剂量升级和延长标记时间对核酸转化RNA测序中的表达估计的影响.
为了确定由广泛的4sU标签引起的表达偏差背后的机制.
开发用于纠正这些偏差的计算和统计方法.

主要方法:

核酸转化RNA测序使用不同度和时间的4sU标记在不同的细胞系进行.
分析的重点是确定与RNA半衰期相关的表达估计中的偏差.
开发一种用于拯救无法映射的读取的计算工具以及用于偏差校正的统计方法.

主要成果:

表达率估计在高4sU度或以后的时间点上以RNA半衰期依赖的方式偏差.
偏差是由于图书馆准备过程中读取可映射性降低,多次转换和标记RNA不足.
一个新的计算工具改善了读取映射,一个统计方法有效地消除了剩余的偏差.

结论:

广泛的4-thiouridine标签可以在RNA测序表达估计中引入显著的偏差.
已开发的计算和统计方法,集成到GRAND-SLAM管道和grandR包中,可以准确地纠正这些偏差.
这些进展使得使用核酸转换RNA-seq. 的新合成RNA能够更可靠地量化.