Advanced Methods in DNA Replication

TOPICAL COLLECTIONS

Guest Editor

Gideon Coster

Institute of Cancer Research, London, UK

Dr Gideon Coster obtained his BSc in Life Sciences from the Hebrew University of Jerusalem in Israel. He carried out his PhD research with Professor Michal Goldberg at the Genetics Department of the Hebrew University of Jerusalem. His work uncovered novel links between the DNA damage response pathway, the cell cycle machinery, and the immune response. Gideon then joined the lab of Dr John Diffley at the Clare Hall Laboratories of the London Research Institute, which later became part of the Francis Crick Institute, to carry out his postdoctoral work. During this time, he uncovered key mechanistic aspects of how the replicative helicase gets loaded onto DNA, work which has far-reaching implications for how our cells initiate the process of bidirectional DNA replication. In October 2018 Gideon joined the Cancer Biology Division of the Institute of Cancer Research (ICR) as a Group Leader, funded by a Sir Henry Dale Fellowship from the Wellcome Trust and the Royal Society. Work in his lab aims to define how difficult to replicate regions of our genome are copied accurately in healthy cells and how this goes wrong in cancer cells.

Collection Overview

An essential requirement for the propagation of life is the faithful transmission of genetic material from one generation to the next. In cellular organisms, this fundamental requirement is fulfilled by a process termed DNA replication. This key process is relevant across multiple fields, ranging from human health and nucleic acid chemistry to enzymology and evolution.

The replisome, a complex piece of machinery, is responsible for carrying out DNA replication. The replisome has to contend with various obstacles, such as DNA-bound proteins and DNA damage. It must coordinate its activity with key cellular pathways, such as transcription and DNA repair. In addition to copying the genetic code, epigenetic information must be restored. These activities are tightly coupled to the cell cycle and coordinated by post-translational modifications (e.g., phosphorylation by the cell cycle and checkpoint kinases).

Recent technological advances have enabled scientists to investigate replication in great detail, including the reconstitution of yeast and human replication in vitro, the high-resolution structures of replisomes and their precursors, and the development of single-molecule methods to visualize the dynamics of individual replication forks. Cellular replication studies use proteomic approaches (iPOND, NCC, and eSPAN) and new techniques like flow cytometry, microscopy, fiber labeling and combing, and sequencing to map replication origins.

This Methods Collection aims to provide cutting-edge methods in DNA replication, covering a wide range of approaches, including biochemistry, single-molecule, microscopy, flow cytometry, sequencing, and proteomics. Together, these detailed protocols will serve as a useful resource for the replication community and beyond.