Study of cell cycle checkpoints using Xenopus cell-free extracts.

Cell cycle checkpoints are involved in the coordinated response to DNA damage and thus play a key role 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 DNA replication, damage, and checkpoint activation. We outline the preparation of cell-free extracts, DNA substrates and their subsequent use in assays aimed at understanding cell cycle checkpoints, and related processes. Several advances made over the years have enabled the continued use of the Xenopus system to answer a variety of questions in DNA replication, repair and checkpoint signaling. It is anticipated that the versatile Xenopus system is amenable to future modification as well to continue studies attempting to understand these important physiological processes.