A Multiplexed System for Quantitative Comparisons of Chromatin Landscapes.
A Multiplexed System for Quantitative Comparisons of Chromatin Landscapes.
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DOI：
10.1016/j.molcel.2015.11.003
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发表时间：
2016-01-07
期刊：
影响因子：
16
通讯作者：
Bernstein BE
中科院分区：
文献类型：
--
作者：
van Galen P;Viny AD;Ram O;Ryan RJ;Cotton MJ;Donohue L;Sievers C;Drier Y;Liau BB;Gillespie SM;Carroll KM;Cross MB;Levine RL;Bernstein BE
组蛋白修饰的全基因组分析可以系统地了解参与给定细胞类型的调控元件和程序。然而，传统的染色质免疫沉淀和测序 (ChIP-seq) 无法捕获组蛋白修饰水平的定量信息，需要大量起始材料，并且涉及对每个样本进行繁琐的处理。在这里，我们通过利用 DNA 条形码以多重格式定量分析染色质的技术来解决这些限制。我们同时绘制多个样本中多个组蛋白修饰的相对水平，每个样本包含少至一千个细胞。我们通过监测 P300、EZH2 或 KDM5 抑制后的动态变化，通过将改变的表观遗传景观与染色质调节突变联系起来，以及通过绘制纯化的人类造血干细胞中的活性和抑制标记来展示该技术。因此，该技术能够对稀有细胞类型、基因型、环境条件和药物治疗的染色质状态动态进行定量研究。
Genome-wide profiling of histone modifications can provide systematic insight into the regulatory elements and programs engaged in a given cell type. However, conventional chromatin immunoprecipitation and sequencing (ChIP-seq) does not capture quantitative information on histone modification levels, requires large amounts of starting material, and involves tedious processing of each individual sample. Here we address these limitations with a technology that leverages DNA barcoding to profile chromatin quantitatively and in multiplexed format. We concurrently map relative levels of multiple histone modifications across multiple samples, each comprising as few as a thousand cells. We demonstrate the technology by monitoring dynamic changes following inhibition of P300, EZH2 or KDM5, by linking altered epigenetic landscapes to chromatin regulator mutations, and by mapping active and repressive marks in purified human hematopoietic stem cells. Hence, this technology enables quantitative studies of chromatin state dynamics across rare cell types, genotypes, environmental conditions and drug treatments.