染色质的特异性分离鉴定了与小鼠胚胎干细胞多能性和体细胞重编程相关的调控因子。

Locus-Specific Isolation of the Chromatin Identifies Regulators Relevant to Pluripotency of Mouse Embryonic Stem Cells and Reprogramming of Somatic Cells.

机构信息

Laboratory of Gene Regulation, Faculty of Medicine, University of Tsukuba, 1-1-1 Tennodai, Tsukuba 305-8575, Japan.

Laboratory of Animal Resource Center, Department of Anatomy and Embryology, Faculty of Medicine, University of Tsukuba, 1-1-1 Tennodai, Tsukuba 305-8575, Japan.

出版信息

Int J Mol Sci. 2022 Dec 3;23(23):15242. doi: 10.3390/ijms232315242.

Abstract

Pluripotency is a crucial feature of pluripotent stem cells, which are regulated by the core pluripotency network consisting of key transcription factors and signaling molecules. However, relatively less is known about the molecular mechanisms that modify the core pluripotency network. Here we used the CAPTURE (CRISPR Affinity Purification in situ of Regulatory Elements) to unbiasedly isolate proteins assembled on the promoter in mouse embryonic stem cells (mESCs), and then tested their functional relevance to the maintenance of mESCs and reprogramming of somatic cells. Gene ontology analysis revealed that the identified proteins, including many RNA-binding proteins (RBPs), are enriched in RNA-related functions and gene expression. ChIP-qPCR experiments confirmed that BCLAF1, FUBP1, MSH6, PARK7, PSIP1, and THRAP3 occupy the promoter region in mESCs. Knockdown experiments of these factors show that they play varying roles in self-renewal, pluripotency gene expression, and differentiation of mESCs as well as in the reprogramming of somatic cells. Our results show the utility of unbiased identification of chromatin-associated proteins on a pluripotency gene in mESCs and reveal the functional relevance of RBPs in ESC differentiation and somatic cell reprogramming.

摘要

多能性是多能干细胞的一个关键特征,多能干细胞受核心多能性网络调控,该网络由关键转录因子和信号分子组成。然而,对于修饰核心多能性网络的分子机制,人们知之甚少。在这里,我们使用 CAPTURE(CRISPR 相关元素的原位亲和纯化)来 unbiasedly 分离在小鼠胚胎干细胞(mESCs)上组装的启动子上的蛋白,然后测试它们对 mESCs 的维持和体细胞重编程的功能相关性。GO 分析显示,所鉴定的蛋白,包括许多 RNA 结合蛋白(RBPs),在 RNA 相关功能和基因表达中富集。ChIP-qPCR 实验证实,BCLAF1、FUBP1、MSH6、PARK7、PSIP1 和 THRAP3 占据 mESCs 中的 启动子区域。这些因子的敲低实验表明,它们在 mESCs 的自我更新、多能性基因表达和分化以及体细胞重编程中发挥不同的作用。我们的结果表明,在 mESCs 中对多能性基因相关染色质蛋白进行无偏鉴定的有效性,并揭示了 RBPs 在 ESC 分化和体细胞重编程中的功能相关性。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/4b65/9740452/9397fde629e8/ijms-23-15242-g001.jpg

文献AI研究员

20分钟写一篇综述,助力文献阅读效率提升50倍

立即体验

用中文搜PubMed

大模型驱动的PubMed中文搜索引擎

马上搜索