A Molecular Toolbox to Engineer Site-Specific DNA Replication Perturbation
Research output: Chapter in Book/Report/Conference proceeding › Book chapter › Research › peer-review
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.
Original language | English |
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Title of host publication | Genome Instability : Methods and Protocols |
Number of pages | 15 |
Volume | 1672 |
Publisher | Springer |
Publication date | 2018 |
Pages | 295-309 |
ISBN (Print) | 978-1-4939-7305-7 |
ISBN (Electronic) | 978-1-4939-7306-4 |
DOIs | |
Publication status | Published - 2018 |
Series | Methods in molecular biology (Clifton, N.J.) |
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ISSN | 1064-3745 |
ID: 190852365