Chromosome conformation capture carbon copy technology.
Chromosome conformation capture carbon copy technology.
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DOI：
10.1002/0471142727.mb2114s80
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发表时间：
2007-10-01
影响因子：
--
通讯作者：
Dekker, Job
中科院分区：
文献类型：
--
作者：
Dostie, Josee;Zhan, Ye;Dekker, Job
染色体构象捕获(3C)用于高分辨率地定量体内的物理DNA接触。在酵母和高等真核生物中，3C最先被用于绘制染色体III的空间染色质组织图，以证明基因组DNA元件通过与靶基因的物理相互作用来调节靶基因。3C已被广泛用于沿着(顺式)或(反式)染色体之间的功能染色质相互作用的小规模分析。对于更大规模的应用，染色体构象捕获碳拷贝(5C)将3C与连接介导的扩增(LMA)相结合，通过微阵列或超高通量DNA测序同时量化数十万个物理DNA接触。5C允许绘制整个基因组中大组基因组元素之间广泛的物理相互作用网络。这类网络可以提供重要的生物学见解，例如，通过确定调控元件与其目标基因之间的关系。本单元描述了用于大规模分析哺乳动物细胞中顺染色质和跨染色质相互作用的5C。
Chromosome conformation capture (3C) is used to quantify physical DNA contacts in vivo at high resolution. 3C was first used in yeast to map the spatial chromatin organization of chromosome III, and in higher eukaryotes to demonstrate that genomic DNA elements regulate target genes by physically interacting with them. 3C has been widely adopted for small-scale analysis of functional chromatin interactions along (cis) or between (trans) chromosomes. For larger-scale applications, chromosome conformation capture carbon copy (5C) combines 3C with ligation-mediated amplification (LMA) to simultaneously quantify hundreds of thousands of physical DNA contacts by microarray or ultra-high-throughput DNA sequencing. 5C allows the mapping of extensive networks of physical interactions among large sets of genomic elements throughout the genome. Such networks can provide important biological insights, e.g., by identifying relationships between regulatory elements and their target genes. This unit describes 5C for large-scale analysis of cis- and trans-chromatin interactions in mammalian cells.