小鼠胚胎成纖維細胞染色質可及性的定量分析

2017年8月，第三軍醫大學大坪醫院創傷、燒傷與復合損傷國家重點實驗室分子生物學中心；中國科學院生物物理研究所生物大分子國家重點實驗室；中國科學院大學；暨南大學寶安婦幼保健院中心實驗室(Molecular Biology Center, State Key Laboratory of Trauma, Burn, and Combined Injury, Daping Hospital, Third Military Medical University, Chongqing 400042, China；National Laboratory for Biomacromolecules, Institute of Biophysics, Chinese Academy of Sciences, Beijing 100101, China；University of Chinese Academy of Sciences, Beijing 100049, China；Central laboratory, Baoan Maternal and Child Health Hospital, Jinan University, Shenzhen 518133, China) Xueqing Xu老師研究團隊在《Biochemical and Biophysical Research Communications》上發表論文：

“Quantitative analysis of chromatin accessibility in mouse embryonic fibroblasts”

“小鼠胚胎成纖維細胞染色質可及性的定量分析”

Abstract：

Genomic DNA of eukaryotic cells is hierarchically packaged into chromatin by histones. The dynamic organization of chromatin fibers plays a critical role in the regulation of gene transcription and other DNA-associated biological processes. Recently, numerous approaches have been developed to map the chromatin organization by characterizing chromatin accessibilities in genome-wide. However, reliable methods to quantitatively map chromatin accessibility are not well-established, especially not on a genome-wide scale. Here, we developed a modified MNase-seq for mouse embryonic fibroblasts, wherein chromatin was partially digested at multiple digestion times using micrococcal nuclease (MNase), allowing quantitative analysis of local yet genome-wide chromatin compaction. Our results provide strong evidence that the chromatin accessibility at promoter regions are positively correlated with gene activity. In conclusion, our assay is an ideal tool for the quantitative study of gene regulation in the perspective of chromatin accessibility.

摘要：

真核細胞的基因組DNA被組蛋白分層包裝成染色質。染色質纖維的動態組織在基因轉錄和其他dna相關生物過程的調控中起著至關重要的作用。最近，已經開發了許多方法，通過表征全基因組染色質可及性來繪制染色質組織。然而，定量繪制染色質可及性的可靠方法尚未建立，特別是在全基因組范圍內。在這里，我們開發了一種用于小鼠胚胎成纖維細胞的改良的MNase-seq，其中使用微球菌核酸酶(MNase)在多次消化時間內部分消化染色質，允許定量分析局部但全基因組的染色質壓實。我們的研究結果提供了強有力的證據，證明啟動子區域的染色質可及性與基因活性呈正相關。總之，從染色質可及性的角度來看，我們的分析是定量研究基因調控的理想工具。

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