Nucleotide-resolution mapping of nascent RNA revisits the principles of transcriptional reorganization of the human genome upon signaling.

新生 RNA 的核苷酸分辨率图谱重新审视了人类基因组根据信号传导进行转录重组的原理。

• 批准号: 290613333
• 负责人: Professor Dr. Argyris Papantonis
• 金额: --
• 依托单位: Abteilung Chromatin-Netzwerke
• 依托单位国家: 德国
• 项目类别: Research Grants
• 财政年份: 2016
• 资助国家: 德国
• 起止时间: 2015-12-31 至 2020-12-31
• 项目状态: 已结题
• 来源: https://gepris.dfg.de/gepris/projekt/290613333?language=en
• 关键词: Nucleotide; resolution; mapping; nascent; RNA

Although human mRNA cataloguing has now become routine, dissecting nascent transcriptomes has been a challenge. The various approaches available to date were, in general, laborious, and focused on just parts of the transcriptome. We recently introduced factory RNA-seq, a methodology that allows the comprehensive capturing of both coding and non-coding nascent RNA at low sequencing depth. However, we still lack understanding on the spatio-temporal deployment of nascent transcription and its connection to RNA processing. With this proposal we aim at further developing our factory RNA-seq approach to achieve single-nucleotide resolution and single-cell trascriptomes, and apply it to human primary endothelial cells before and after cytokine stimulation with two cytokines, TNFalpha and TGFbeta. This way we will be able to address topology and temporal succession in the production of nascent RNAs, assess their involvement in the active sites of transcription, and ultimately revisit our current understanding of mammalian transcriptome reorganization upon signaling at unprecedented resolution.

尽管人类 mRNA 编目现已成为常规，但剖析新生转录组一直是一个挑战。迄今为止可用的各种方法总体来说都是费力的，并且只关注转录组的一部分。我们最近推出了工厂 RNA-seq，这是一种能够以低测序深度全面捕获编码和非编码新生 RNA 的方法。然而，我们仍然缺乏对新生转录的时空部署及其与 RNA 加工的联系的了解。通过这项提案，我们的目标是进一步开发我们的工厂 RNA-seq 方法，以实现单核苷酸分辨率和单细胞转录组，并将其应用于用两种细胞因子 TNFα 和 TGFβ 刺激之前和之后的人原代内皮细胞。通过这种方式，我们将能够解决新生RNA产生过程中的拓扑结构和时间连续性问题，评估它们在转录活性位点中的参与，并最终以前所未有的分辨率重新审视我们目前对哺乳动物转录组重组的理解。