Analyzing genome integrity during DNA replication using iPOND (isolation of Proteins On Nascent DNA)

Abstract

The in vivo analysis of dynamic events that protect genomic and epigenomic integrity during DNA replication has been a challenging task. The proteins that ensure the accurate inheritance of the genome act directly at replication forks. Unfortunately, tracking and purifying replication forks has been difficult in mammalian cells because the location of elongating forks is difficult to predict. I have developed the iPOND (isolation of Proteins On Nascent DNA) technique as a tool to isolate, analyze and discover proteins that localize to replication forks during active DNA replication and following conditions of DNA damage. The iPOND technique provides a useful tool to investigate the coordinated mechanisms that maintain the genome and epigenome through DNA replication. These pathways are perturbed in human diseases such as cancer.