Abstract
Ataxia Telangiectasia (A-T) is caused by biallelic inactivation of the Ataxia Telangiectasia Mutated (ATM) gene, due to nonsense or missense mutations, small insertions/deletions (indels), splicing alterations, and large genomic rearrangements. After establishing A-T clinical diagnosis, a molecular confirmation is needed, based on the detection of one of these loss-of-function mutations in at least one allele. In most cases, the pathogenicity of the detected mutations is sufficient to make a definitive diagnosis. More rarely, mutations of unknown consequences are identified and direct biological analyses are required to establish their pathogenic characters. In such cases, complementary analyses of ATM expression, localization, and activity allow fine characterization of these mutations and facilitate A-T diagnosis. Here, we present genetic and biochemical protocols currently used in the laboratory that have proven to be highly accurate, reproducible, and quantitative. We also provide additional discussion on the critical points of the techniques presented here.
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Acknowledgments
G.R. is supported by grants from Ligue Nationale Contre le Cancer and Fondation pour la Recherche Médicale n°FDT20130928274. This work was supported in part by a grant of the Fondation AT EUROPE. We thank Genethon for access to cell lines, and the Institut Curie sequencing facility.
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Rieunier, G., D’Enghien, C.D., Fievet, A., Bellanger, D., Stoppa-Lyonnet, D., Stern, MH. (2017). ATM Gene Mutation Detection Techniques and Functional Analysis. In: Kozlov, S. (eds) ATM Kinase. Methods in Molecular Biology, vol 1599. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-6955-5_3
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DOI: https://doi.org/10.1007/978-1-4939-6955-5_3
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