Native elongating transcript sequencing (NET-seq)

2023-10-10 16:18:07 星期二

Net-seq（原位延伸转录测序）是一种用于以核苷酸分辨率跨基因组分析RNA聚合酶（Pol）活性的方法。以下是Net-seq的简要概述：

目的: Net-seq旨在捕获并测序仍与RNA聚合酶结合在一起的活跃转录过程中的RNA的3'末端。这有助于创建跨基因组的活跃转录位点的高分辨率图。

工作原理:

• 从细胞中提取RNA聚合酶及其绑定的RNA。
• 使用特定的方法仅选择与RNA聚合酶结合的RNA的3'端。
• 对这些3'端进行测序，从而获得在转录过程中的RNA的精确位置。

这种技术为研究基因的精确转录动态提供了一个强大的工具。

BRD4, or Bromodomain-containing protein 4, is a member of the bromodomain and extraterminal (BET) family of proteins. Here's a brief overview:

Function: BRD4 is involved in several crucial cellular processes:

• Transcriptional Regulation: BRD4 plays a vital role in promoting the transcription of many genes by associating with acetylated chromatin. It's particularly significant in the regulation of genes associated with the cell cycle and can recruit positive transcription elongation factor b (P-TEFb) to promote transcriptional elongation.
• Cell Cycle Progression: BRD4 is essential for progressing through various cell cycle stages, especially the G1/S transition.

Medical Relevance: Given its role in transcription and cell cycle regulation, it's not surprising that BRD4 has been implicated in cancer biology. Inhibitors targeting the bromodomains of BRD4, commonly known as BET inhibitors, are being explored as potential therapeutic agents for various cancers.

Structure: BRD4 contains two bromodomains. Bromodomains can recognize and bind acetylated lysine residues, which are common post-translational modifications on histones and other proteins.

Associated Diseases: Alterations, mutations, or aberrant expression of BRD4 has been implicated in several cancers, including acute myeloid leukemia (AML), midline carcinoma, and others.

Inhibitors: Several small molecules that inhibit BRD4 have been developed as potential therapeutic agents. Notably, drugs like JQ1 and I-BET762 bind to the bromodomains of BRD4, inhibiting its interaction with acetylated proteins.

BRD4 is an active area of research, and new findings related to its function, regulation, and potential as a therapeutic target are regularly published.

HiS-NET-seq profiles the active enhancer landscape Putative enhancers show enhancer activity HiS-NET-seq reveals 11,007 new transcribed putative enhancers in K562 cells providing a more complete view on active enhancers. ChIP-Rx vs HiS-NET-seq: BRD4 is required for genome-wide enhancer transcription. HiChIP (H3K27Ac) is for discovering Enhancer connecting Promoter Enhancer and traget gene transcription is regulated by BRD4 Identify other post-initiation transcription regulators ablation: 切除,风化,消融,除去,切除 Acute BRD4-loss impairs transcription at the 3'-end genes. Acute BRD4-loss impairs transcription at the 5' and 3'-end genes. BET & BRD4 degradation prompt (激励,鼓动,提示) 3'-processing defects Readthrough gene How do the distinct molecular interactions mediated by BRD4's bromodomains contribute to its diverse roles in cellular processes, and how can this knowledge be leveraged to develop more targeted and effective therapeutic strategies for BRD4-associated diseases?

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