FORK-seq: replication landscape of the Saccharomyces cerevisiae genome by nanopore sequencing
FORK-seq: replication landscape of the Saccharomyces cerevisiae genome by nanopore sequencing
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DOI：
10.1186/s13059-020-02013-3
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发表时间：
2020-05-26
期刊：
影响因子：
12.3
通讯作者：
Hyrien, Olivier
中科院分区：
文献类型：
--
作者：
Hennion, Magali;Arbona, Jean-Michel;Hyrien, Olivier
基因组复制作图方法描绘细胞群体，掩盖细胞间异质性。在这里，我们描述了FORK-seq，一种纳米孔测序方法，以200个核苷酸的分辨率映射单个DNA分子的复制。通过定量BrdU掺入沿着脉冲追逐复制中间体从酿酒酵母，我们定向58,651复制轨道再现基于群体的复制方向性配置文件和地图4964和4485个人的起始和终止事件，分别。虽然大多数事件聚集在已知的起源和分叉合并区，但9%和18%的启动和终止事件分别发生在以前错过的许多位置。因此，FORK-seq揭示了DNA复制中细胞间异质性的全部程度。
Genome replication mapping methods profile cell populations, masking cell-to-cell heterogeneity. Here, we describe FORK-seq, a nanopore sequencing method to map replication of single DNA molecules at 200-nucleotide resolution. By quantifying BrdU incorporation along pulse-chased replication intermediates from Saccharomyces cerevisiae, we orient 58,651 replication tracks reproducing population-based replication directionality profiles and map 4964 and 4485 individual initiation and termination events, respectively. Although most events cluster at known origins and fork merging zones, 9% and 18% of initiation and termination events, respectively, occur at many locations previously missed. Thus, FORK-seq reveals the full extent of cell-to-cell heterogeneity in DNA replication.