The comet assay is a versatile method to detect nuclear DNA damage in individual eukaryotic cells, from yeast to human. The types of damage detected encompass DNA strand breaks and alkali-labile sites (e.g., apurinic/apyrimidinic sites), alkylated and oxidized nucleobases, DNA–DNA crosslinks, UV-induced cyclobutane pyrimidine dimers and some chemically induced DNA adducts. Depending on the specimen type, there are important modifications to the comet assay protocol to avoid the formation of additional DNA damage during the processing of samples and to ensure sufficient sensitivity to detect differences in damage levels between sample groups. Various applications of the comet assay have been validated by research groups in academia, industry and regulatory agencies, and its strengths are highlighted by the adoption of the comet assay as an in vivo test for genotoxicity in animal organs by the Organisation for Economic Co-operation and Development. The present document includes a series of consensus protocols that describe the application of the comet assay to a wide variety of cell types, species and types of DNA damage, thereby demonstrating its versatility.

Measuring DNA modifications with the comet assay: a compendium of protocols / C. Andrew, M. Peter, G. Goran, V. Soňa, A. Abdulhadi, A. Diana, Ezgi Eyluel Bankoglu, B. Stefano, B. Elisa, B. Gunnar, C. Christy, M.S. Cooke, C. Carla, C. Solange, D. Alok, Joaquin de Lapuente, C. DEL BO', D. Julien, D. Maria, S.J. Duthie, Naouale El Yamani, E. Bevin, G. Isabel, G. Lisa, G. Roger, G. Sofia, K.B. Gutzkow, H. Khaled, H. Alba, H. Oscar, I. Marina, A.N. Jha, K. Siegfried, I.M. Kooter, K. Gudrun, K. Marcin, L. Carina, L. Blanca, L. Marcelo, Ludovic Le Hégarat, L. Angélique, L. Anna, G.E. Liwszyc, Adela López de Cerain, M. Mugimane, M. Ricard, M. Mirta, Vanessa Moraes de Andrade, M. Massimo, M. Damian, N. Matjaž, O. Rui, O. Ann-Karin, O. Norah, P. Mário, A.K. Pandey, P. Stefan, P. Bertrand, R. Kerstin, R. Emilio, R. Elise, S. Julen, S. Sergey, S. Ville, S. Karen, S. Helga, João Paulo Teixeira, V. Vanessa, V. Mahara, Frederique van Acker, Frederik-Jan van Schooten, V. Marie, J.F. Wentzel, W. Maciej, W. Annelies, Ž. Bojana, Z. Tomas, S.A.S. Langie &, A. Amaya. - In: NATURE PROTOCOLS. - ISSN 1754-2189. - (2023). [Epub ahead of print] [10.1038/s41596-022-00754-y]

Measuring DNA modifications with the comet assay: a compendium of protocols

C. DEL BO';
2023

Abstract

The comet assay is a versatile method to detect nuclear DNA damage in individual eukaryotic cells, from yeast to human. The types of damage detected encompass DNA strand breaks and alkali-labile sites (e.g., apurinic/apyrimidinic sites), alkylated and oxidized nucleobases, DNA–DNA crosslinks, UV-induced cyclobutane pyrimidine dimers and some chemically induced DNA adducts. Depending on the specimen type, there are important modifications to the comet assay protocol to avoid the formation of additional DNA damage during the processing of samples and to ensure sufficient sensitivity to detect differences in damage levels between sample groups. Various applications of the comet assay have been validated by research groups in academia, industry and regulatory agencies, and its strengths are highlighted by the adoption of the comet assay as an in vivo test for genotoxicity in animal organs by the Organisation for Economic Co-operation and Development. The present document includes a series of consensus protocols that describe the application of the comet assay to a wide variety of cell types, species and types of DNA damage, thereby demonstrating its versatility.
Settore MED/49 - Scienze Tecniche Dietetiche Applicate
2023
27-gen-2023
Article (author)
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Utilizza questo identificativo per citare o creare un link a questo documento: https://hdl.handle.net/2434/953215
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