Microscopy-Based Chromosome Conformation Capture Enables Simultaneous Visualization of Genome Organization and Transcription in Intact Organisms
Microscopy-Based Chromosome Conformation Capture Enables Simultaneous Visualization of Genome Organization and Transcription in Intact Organisms
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DOI：
10.1016/j.molcel.2019.01.011
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发表时间：
2019-04-04
期刊：
影响因子：
16
通讯作者：
Nollmann, Marcelo
中科院分区：
文献类型：
--
作者：
Cardozo Gizzi, Andres M.;Cattoni, Diego, I;Nollmann, Marcelo
真核细胞染色体以多种尺度组织，从核小体到染色体区域。最近，全基因组的方法确定了一个中间水平的染色体组织，拓扑关联结构域（TADS），在转录调控中发挥关键作用。然而，这些方法不能直接检查转录激活和染色体结构之间的相互作用，同时保持空间信息。在这里，我们提出了一个多路复用，顺序成像方法（Hi-M），允许在单个细胞核中的染色体组织和转录的同时检测。这使我们能够揭示在完整的果蝇胚胎中合子基因组唤醒期间转录激活和同源染色体解配对后发生的3D染色质组织的变化。令人兴奋的是，Hi-M在不同发育阶段或细胞周期中探索具有空间分辨率的多尺度染色体结构的能力将是理解基因组4D组织的机制和后果的关键。
Eukaryotic chromosomes are organized in multiple scales, from nucleosomes to chromosome territories. Recently, genome-wide methods identified an intermediate level of chromosome organization, topologically associating domains (TADS), that play key roles in transcriptional regulation. However, these methods cannot directly examine the interplay between transcriptional activation and chromosome architecture while maintaining spatial information. Here we present a multiplexed, sequential imaging approach (Hi-M) that permits simultaneous detection of chromosome organization and transcription in single nuclei. This allowed us to unveil the changes in 3D chromatin organization occurring upon transcriptional activation and homologous chromosome unpairing during awakening of the zygotic genome in intact Drosophila embryos. Excitingly, the ability of Hi-M to explore the multi-scale chromosome architecture with spatial resolution at different stages of development or during the cell cycle will be key to understanding the mechanisms and consequences of the 4D organization of the genome.