小鼠胚胎成纤维细胞染色质可及性的定量分析

2017年8月，第三军医大学大坪医院创伤、烧伤与复合损伤国家重点实验室分子生物学中心；中国科学院生物物理研究所生物大分子国家重点实验室；中国科学院大学；暨南大学宝安妇幼保健院中心实验室(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老师研究团队在《Biochemical and Biophysical Research Communications》上发表论文：

“Quantitative analysis of chromatin accessibility in mouse embryonic fibroblasts”

“小鼠胚胎成纤维细胞染色质可及性的定量分析”

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：

真核细胞的基因组DNA被组蛋白分层包装成染色质。染色质纤维的动态组织在基因转录和其他dna相关生物过程的调控中起着至关重要的作用。最近，已经开发了许多方法，通过表征全基因组染色质可及性来绘制染色质组织。然而，定量绘制染色质可及性的可靠方法尚未建立，特别是在全基因组范围内。在这里，我们开发了一种用于小鼠胚胎成纤维细胞的改良的MNase-seq，其中使用微球菌核酸酶(MNase)在多次消化时间内部分消化染色质，允许定量分析局部但全基因组的染色质压实。我们的研究结果提供了强有力的证据，证明启动子区域的染色质可及性与基因活性呈正相关。总之，从染色质可及性的角度来看，我们的分析是定量研究基因调控的理想工具。

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