A Molecular Toolbox to Engineer Site-Specific DNA Replication Perturbation

Nicolai B Larsen, Ian D Hickson, Hocine W Mankouri

4 Citationer (Scopus)

Abstract

Site-specific arrest of DNA replication is a useful tool for analyzing cellular responses to DNA replication perturbation. The E. coli Tus-Ter replication barrier can be reconstituted in eukaryotic cells as a system to engineer an unscheduled collision between a replication fork and an "alien" impediment to DNA replication. To further develop this system as a versatile tool, we describe a set of reagents and a detailed protocol that can be used to engineer Tus-Ter barriers into any locus in the budding yeast genome. Because the Tus-Ter complex is a bipartite system with intrinsic DNA replication-blocking activity, the reagents and protocols developed and validated in yeast could also be optimized to engineer site-specific replication fork barriers into other eukaryotic cell types.

OriginalsprogEngelsk
TitelGenome Instability : Methods and Protocols
Antal sider15
Vol/bind1672
ForlagSpringer
Publikationsdato2018
Sider295-309
ISBN (Trykt)978-1-4939-7305-7
ISBN (Elektronisk)978-1-4939-7306-4
DOI
StatusUdgivet - 2018
NavnMethods in molecular biology (Clifton, N.J.)
ISSN1064-3745

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