Single-Cell DamID to Capture Contacts Between DNA and the Nuclear Lamina in Individual Mammalian Cells
Single-Cell DamID to Capture Contacts Between DNA and the Nuclear Lamina in Individual Mammalian Cells
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DOI：
10.1007/978-1-0716-0664-3_9
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发表时间：
2021-01-01
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影响因子：
--
通讯作者：
Kind, Jop
中科院分区：
文献类型：
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作者：
De Luca, Kim L.;Kind, Jop
真核细胞内DNA的组织对于基因表达的调节和DNA损伤的修复等细胞过程是重要的。为了理解复杂系统中细胞间的差异，对单个细胞进行系统分析是必要的。虽然有许多工具可以捕获DNA构象和染色质背景，但这些方法通常需要大量的细胞才能获得足够的输出。在这里，我们描述了单细胞DamID，这是一种捕捉DNA和特定感兴趣蛋白质之间的接触的技术。通过将细菌甲基转移酶Dam与核层蛋白lamin B1融合，可以绘制与核周接触的基因组区域。单细胞DAMID生成具有足够吞吐量和分辨率的接触图，以可靠地识别间期染色体核组织的相似和变异模式。
The organization of DNA within the eukaryotic nucleus is important for cellular processes such as regulation of gene expression and repair of DNA damage. To comprehend cell-to-cell variation within a complex system, systematic analysis of individual cells is necessary. While many tools exist to capture DNA conformation and chromatin context, these methods generally require large populations of cells for sufficient output. Here we describe single-cell DamID, a technique to capture contacts between DNA and a given protein of interest. By fusing the bacterial methyltransferase Dam to nuclear lamina protein lamin B1, genomic regions in contact with the nuclear periphery can be mapped. Single-cell DamID generates contact maps with sufficient throughput and resolution to reliably identify patterns of similarity as well as variation in nuclear organization of interphase chromosomes.