High-throughput analysis of single human cells reveals the complex nature of DNA replication timing control.
High-throughput analysis of single human cells reveals the complex nature of DNA replication timing control.
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DOI：
10.1038/s41467-022-30212-y
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发表时间：
2022-05-03
影响因子：
16.6
通讯作者：
中科院分区：
文献类型：
--
作者：
DNA复制从复制起始点开始，贯穿整个S期。关于起源是一组固定的基因座，还是在单个细胞中随机使用的潜在位点的松散聚集，以及它们的放电时间有多一致，争论仍然存在。我们开发了一种从数千个单细胞的全基因组测序中分析DNA复制的方法，其中包括硅流式细胞术，一种区分复制和非复制细胞的方法。使用两个微流控平台，我们分析多达2437个复制细胞从一个单一的样本。数据的分辨率和规模允许集中分析复制起始位点，表明大多数发生在有限的基因组区域。虽然起始顺序在细胞间非常相似，但我们意外地在后期复制区域发现了几个亚型的起始区域。总之，单个细胞的高通量、高分辨率测序揭示了以前被低估的复制起始和进展的可变性。在人类细胞系中，DNA复制主要是从单个细胞中共享的一组基因组位点开始的，这些基因组位点根据可预测但不是严格确定的顺序进行。
DNA replication initiates from replication origins firing throughout S phase. Debate remains about whether origins are a fixed set of loci, or a loose agglomeration of potential sites used stochastically in individual cells, and about how consistent their firing time is. We develop an approach to profile DNA replication from whole-genome sequencing of thousands of single cells, which includes in silico flow cytometry, a method for discriminating replicating and non-replicating cells. Using two microfluidic platforms, we analyze up to 2437 replicating cells from a single sample. The resolution and scale of the data allow focused analysis of replication initiation sites, demonstrating that most occur in confined genomic regions. While initiation order is remarkably similar across cells, we unexpectedly identify several subtypes of initiation regions in late-replicating regions. Taken together, high throughput, high resolution sequencing of individual cells reveals previously underappreciated variability in replication initiation and progression. In human cell lines, DNA replication initiates primarily from a set of genomic loci shared across single cells, which fire according to a predictable but not strictly deterministic order.
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影响因子：
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作者：
Langley AR;Gräf S;Smith JC;Krude T
通讯作者：
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