Telomere-to-telomere DNA replication timing profiling using single-molecule sequencing with Nanotiming.

ABSTRACT: Current temporal studies of DNA replication are either low-resolution or require complex cell synchronisation and/or sorting procedures. Here we introduce Nanotiming, a single-molecule, nanopore sequencing-based method producing high-resolution, telomere-to-telomere replication timing (RT) profiles of eukaryotic genomes by interrogating changes in intracellular dTTP concentration during S phase through competition with its analogue bromodeoxyuridine triphosphate (BrdUTP) for incorporation into replicating DNA. This solely demands the labelling of asynchronously growing cells with an innocuous dose of BrdU during one doubling time followed by BrdU quantification along nanopore reads. We demonstrate in S. cerevisiae model eukaryote that Nanotiming reproduces RT profiles generated by referenc

SUBMITTER: Theulot B 

PROVIDER: S-EPMC11696806 | biostudies-literature | 2025 Jan

REPOSITORIES: biostudies-literature