High-resolution mapping of mitotic DNA synthesis under conditions of replication stress in cultured cells

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 aphidic...

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Autores principales: Groelly, Florian J., Dagg, Rebecca A., Mailler, Jonathan, Halazonetis, Thanos D., Tarsounas, Madalena
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Elsevier 2023
Materias:
Protocol
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9826876/
https://www.ncbi.nlm.nih.gov/pubmed/36598851
http://dx.doi.org/10.1016/j.xpro.2022.101970
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author Groelly, Florian J.
Dagg, Rebecca A.
Mailler, Jonathan
Halazonetis, Thanos D.
Tarsounas, Madalena
author_facet Groelly, Florian J.
Dagg, Rebecca A.
Mailler, Jonathan
Halazonetis, Thanos D.
Tarsounas, Madalena
author_sort Groelly, Florian J.
collection PubMed
description 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)
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spelling pubmed-98268762023-01-10 High-resolution mapping of mitotic DNA synthesis under conditions of replication stress in cultured cells Groelly, Florian J. Dagg, Rebecca A. Mailler, Jonathan Halazonetis, Thanos D. Tarsounas, Madalena STAR Protoc Protocol 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) Elsevier 2023-01-03 /pmc/articles/PMC9826876/ /pubmed/36598851 http://dx.doi.org/10.1016/j.xpro.2022.101970 Text en © 2022. https://creativecommons.org/licenses/by-nc-nd/4.0/This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/).
spellingShingle Protocol
Groelly, Florian J.
Dagg, Rebecca A.
Mailler, Jonathan
Halazonetis, Thanos D.
Tarsounas, Madalena
High-resolution mapping of mitotic DNA synthesis under conditions of replication stress in cultured cells
title High-resolution mapping of mitotic DNA synthesis under conditions of replication stress in cultured cells
title_full High-resolution mapping of mitotic DNA synthesis under conditions of replication stress in cultured cells
title_fullStr High-resolution mapping of mitotic DNA synthesis under conditions of replication stress in cultured cells
title_full_unstemmed High-resolution mapping of mitotic DNA synthesis under conditions of replication stress in cultured cells
title_short High-resolution mapping of mitotic DNA synthesis under conditions of replication stress in cultured cells
title_sort high-resolution mapping of mitotic dna synthesis under conditions of replication stress in cultured cells
topic Protocol
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9826876/
https://www.ncbi.nlm.nih.gov/pubmed/36598851
http://dx.doi.org/10.1016/j.xpro.2022.101970
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