Multiple laser lines that are used for single-molecule TIRF (total-internal fluorescence) imaging

Introduction

Complete and faithful duplication of the genome is crucial to support proper cell division and maintain the genetic integrity of all organisms. In particular, errors in DNA replication can lead to genomic instability, a characteristic feature of most cancer cells.

A simplified cartoon depiction of the eukaryotic replisome complex.

A simplified cartoon depiction of the eukaryotic replisome complex.

Our laboratory aims to study eukaryotic DNA replication at the single-molecule level that will lend important new insights into the mechanism and dynamics of the complex molecular machines that carry out genome duplication.

A major advantage of single-molecule methods is that measurements are made on individual molecules, allowing their kinetics to be determined directly. Therefore, one can gain essential information about transient dynamics and heterogeneities which are not accessible using conventional biochemical approaches. In addition, recent advances in nano-manipulation methods allow measuring the physical properties of molecules in response to external forces.

We use a number of model systems including extracts derived from Xenopus laevis eggs and S.cerevisiae as well as the Simian Virus 40 (SV40) replication system.