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Cell Cycle Checkpoints pp 103–144Cite as

DNA Damage Response in Xenopus laevis Cell-Free Extracts

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Part of the book series: Methods in Molecular Biology ((MIMB,volume 2267))

Abstract

The DNA damage response (DDR) is a coordinated cellular response to a variety of insults to the genome. DDR initiates the activation of cell cycle checkpoints preventing the propagation of damaged DNA followed by DNA repair, which are both critical in maintaining genome integrity. Several model systems have been developed to study the mechanisms and complexity of checkpoint function. Here we describe the application of cell-free extracts derived from Xenopus eggs as a model system to investigate signaling from DNA damage, modulation of DNA replication, checkpoint activation, and ultimately DNA repair. We outline the preparation of cell-free extracts, DNA substrates, and their subsequent use in assays aimed at understanding the cellular response to DNA damage. Cell-free extracts derived from the eggs of Xenopus laevis remain a robust and versatile system to decipher the biochemical steps underlying this essential characteristic of all cells, critical for genome stability.

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Acknowledgments

This work was supported by grants from the National Institutes of Health P01 CA174653 and R35 CA197606 to J.G. and R50 CA233182 to T.A.

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Authors and Affiliations

  1. Institute for Cancer Genetics, College of Physicians and Surgeons, Columbia University, New York, USA

    Tomas Aparicio Casado & Jean Gautier

  2. Department of Genetics and Development, Columbia University School of Medicine, New York, USA

    Jean Gautier

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  1. Tomas Aparicio Casado
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  2. Jean Gautier
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Correspondence to Jean Gautier .

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Editors and Affiliations

  1. Department of Oncological Sciences, Icahn School of Medicine at Mount Sinai, New York, NY, USA

    James J. Manfredi

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Aparicio Casado, T., Gautier, J. (2021). DNA Damage Response in Xenopus laevis Cell-Free Extracts. In: Manfredi, J.J. (eds) Cell Cycle Checkpoints. Methods in Molecular Biology, vol 2267. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-1217-0_8

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  • DOI: https://doi.org/10.1007/978-1-0716-1217-0_8

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