A Live-Cell Imaging Approach for Measuring DNA Replication Rates

Daniel Dovrat, Danielle Dahan, Shachar Sherman, Ioannis Tsirkas, Natalie Elia, Amir Aharoni

Research output: Contribution to journalArticlepeer-review

10 Scopus citations

Abstract

We describe a simple and direct approach to measure the progression of single DNA replication forks in living cells by monitoring two fluorescently labeled loci downstream of an origin of replication. We employ this approach to investigate the roles of several leading and lagging strand factors in overall replisome function and show that fork progression is strongly dependent on proper maturation of Okazaki fragments. We also demonstrate how related cellular phenotypes, such as cell-cycle progression and the dynamics of sister chromatid cohesion, may be simultaneously monitored and correlated to DNA replication at the single-cell level.

Original languageEnglish
Pages (from-to)252-258
Number of pages7
JournalCell Reports
Volume24
Issue number1
DOIs
StatePublished - 3 Jul 2018

Keywords

  • DNA replication
  • FROS
  • Okazaki fragment maturation
  • live-cell imaging
  • sister chromatid cohesion

ASJC Scopus subject areas

  • Biochemistry, Genetics and Molecular Biology (all)

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