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Kronos scRT: a uniform framework for single-cell replication timing analysis

By Stefano Gnan, Joseph M. Josephides, Xia Wu, Manuela Spagnuolo, Dalila Saulebekova, Myl&egravene Bohec, Marie Dumont, Laura G Baudrin, Daniele Fachinetti, Sylvain Baulande, Chun-Long Chen

Posted 03 Sep 2021
bioRxiv DOI: 10.1101/2021.09.01.458599

Mammalian genomes are replicated in a cell-type specific order and in coordination with transcription and chromatin organization. Although the field of replication is also entering the single-cell era, current studies require cell sorting, individual cell processing and have yielded a limited number (<100) of cells. Here, we have developed Kronos scRT (https://github.com/CL-CHEN-Lab/Kronos_scRT), a software for single-cell Replication Timing (scRT) analysis. Kronos scRT does not require a specific platform nor cell sorting, allowing the investigation of large datasets obtained from asynchronous cells. Analysis of published available data and droplet-based scWGS data generated in the current study, allows exploitation of scRT data from thousands of cells for different mouse and human cell lines. Our results demonstrate that, although most cells replicate within a close timing range for a given genomic region, replication can also occur stochastically throughout S phase. Altogether, Kronos scRT allows investigating the RT program at a single-cell resolution for both homogeneous and heterogeneous cell populations in a fast and comprehensive manner.

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