High-resolution mapping of mitotic DNA synthesis under conditions of replication stress in cultured cells.
| Autor: | Groelly FJ; Genome Stability and Tumourigenesis Group, Department of Oncology, Oxford Institute for Radiation Oncology, University of Oxford, Oxford OX3 7DQ, UK., Dagg RA; Genome Stability and Tumourigenesis Group, Department of Oncology, Oxford Institute for Radiation Oncology, University of Oxford, Oxford OX3 7DQ, UK., Mailler J; Department of Molecular Biology, University of Geneva, 1205 Geneva, Switzerland., Halazonetis TD; Department of Molecular Biology, University of Geneva, 1205 Geneva, Switzerland. Electronic address: thanos.halazonetis@unige.ch., Tarsounas M; Genome Stability and Tumourigenesis Group, Department of Oncology, Oxford Institute for Radiation Oncology, University of Oxford, Oxford OX3 7DQ, UK. Electronic address: madalena.tarsounas@oncology.ox.ac.uk. |
|---|---|
| Jazyk: | angličtina |
| Zdroj: | STAR protocols [STAR Protoc] 2023 Mar 17; Vol. 4 (1), pp. 101970. Date of Electronic Publication: 2023 Jan 03. |
| DOI: | 10.1016/j.xpro.2022.101970 |
| Abstrakt: | Cells experiencing DNA replication stress enter mitosis with under-replicated DNA, which activates a repair mechanism known as mitotic DNA synthesis (MiDAS). Here we describe a protocol to identify at genome wide and at high resolution the genomic sites where MiDAS occurs in cells exposed to aphidicolin. We use EdU incorporation to label nascent DNA in mitotic cells, followed by isolation of the EdU-labeled DNA and next-generation sequencing. For complete details on the use and execution of this protocol, please refer to Groelly et al. (2022) 1 and Macheret et al. (2020). 2 . Competing Interests: Declaration of interests T.D.H. has a part-time position as Chief Scientific Officer of FoRx Therapeutics, AG. (Copyright © 2023 The Authors. Published by Elsevier Inc. All rights reserved.) |
| Databáze: | MEDLINE |
| Externí odkaz: |
|