1. Measuring DNA modifications with the comet assay: a compendium of protocols
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Andrew Collins, Peter Møller, Goran Gajski, Soňa Vodenková, Abdulhadi Abdulwahed, Diana Anderson, Ezgi Eyluel Bankoglu, Stefano Bonassi, Elisa Boutet-Robinet, Gunnar Brunborg, Christy Chao, Marcus S. Cooke, Carla Costa, Solange Costa, Alok Dhawan, Joaquin de Lapuente, Cristian Del Bo’, Julien Dubus, Maria Dusinska, Susan J. Duthie, Naouale El Yamani, Bevin Engelward, Isabel Gaivão, Lisa Giovannelli, Roger Godschalk, Sofia Guilherme, Kristine B. Gutzkow, Khaled Habas, Alba Hernández, Oscar Herrero, Marina Isidori, Awadhesh N. Jha, Siegfried Knasmüller, Ingeborg M. Kooter, Gudrun Koppen, Marcin Kruszewski, Carina Ladeira, Blanca Laffon, Marcelo Larramendy, Ludovic Le Hégarat, Angélique Lewies, Anna Lewinska, Guillermo E. Liwszyc, Adela López de Cerain, Mugimane Manjanatha, Ricard Marcos, Mirta Milić, Vanessa Moraes de Andrade, Massimo Moretti, Damian Muruzabal, Matjaž Novak, Rui Oliveira, Ann-Karin Olsen, Norah Owiti, Mário Pacheco, Alok K. Pandey, Stefan Pfuhler, Bertrand Pourrut, Kerstin Reisinger, Emilio Rojas, Elise Rundén-Pran, Julen Sanz-Serrano, Sergey Shaposhnikov, Ville Sipinen, Karen Smeets, Helga Stopper, João Paulo Teixeira, Vanessa Valdiglesias, Mahara Valverde, Frederique van Acker, Frederik-Jan van Schooten, Marie Vasquez, Johannes F. Wentzel, Maciej Wnuk, Annelies Wouters, Bojana Žegura, Tomas Zikmund, Sabine A. S. Langie, Amaya Azqueta, University of Oslo (UiO), University of Copenhagen = Københavns Universitet (UCPH), Institute for Medical Research and Occupational Health, Institute of Experimental Medicine of the Academy of Sciences of the Czech Republic, Charles University [Prague] (CU), Florida International University [Miami] (FIU), University of Bradford, University of Würzburg = Universität Würzburg, Istituto di Ricovero e Cura a Carattere Scientifico (IRCCS San Raffaele Pisana), Universita Vita Salute San Raffaele = Vita-Salute San Raffaele University [Milan, Italie] (UniSR), ToxAlim (ToxAlim), Université Toulouse III - Paul Sabatier (UT3), Université de Toulouse (UT)-Université de Toulouse (UT)-Ecole Nationale Vétérinaire de Toulouse (ENVT), Institut National Polytechnique (Toulouse) (Toulouse INP), Université de Toulouse (UT)-Université de Toulouse (UT)-Institut National Polytechnique (Toulouse) (Toulouse INP), Université de Toulouse (UT)-Ecole d'Ingénieurs de Purpan (INP - PURPAN), Université de Toulouse (UT)-Université de Toulouse (UT)-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE), Centre for Environmental Radioactivity (CERAD CoE), Norvegian Institut of Public Health, Massachusetts Institute of Technology (MIT), University of South Florida [Tampa] (USF), Instituto Nacional de Saùde Dr Ricardo Jorge [Portugal] (INSA), Universidade do Porto = University of Porto, Centre for Biomedical Research, University of Hull, University of Hull, Institut de Biosciences et Biotechnologies d'Aix-Marseille (ex-IBEB) (BIAM), Aix Marseille Université (AMU)-Centre National de la Recherche Scientifique (CNRS)-Direction de Recherche Fondamentale (CEA) (DRF (CEA)), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA), Service d'Etudes du Comportement des Radionucléides (SECR), Département de Physico-Chimie (DPC), CEA-Direction des Energies (ex-Direction de l'Energie Nucléaire) (CEA-DES (ex-DEN)), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Paris-Saclay-CEA-Direction des Energies (ex-Direction de l'Energie Nucléaire) (CEA-DES (ex-DEN)), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Paris-Saclay, Scientific Committee for Consumer Safety (SCCS) (SCCS), University of Aberdeen, Laboratoire de Fougères - ANSES, Agence nationale de sécurité sanitaire de l'alimentation, de l'environnement et du travail (ANSES), Farmacologie en Toxicologie, RS: NUTRIM - R3 - Respiratory & Age-related Health, Collins, Andrew, Møller, Peter, Gajski, Goran, Vodenková, Soňa, Abdulwahed, Abdulhadi, Anderson, Diana, Bankoglu, Ezgi Eyluel, Bonassi, Stefano, Boutet-Robinet, Elisa, Brunborg, Gunnar, Chao, Christy, Cooke, Marcus S, Costa, Carla, Costa, Solange, Dhawan, Alok, de Lapuente, Joaquin, Bo', Cristian Del, Dubus, Julien, Dusinska, Maria, Duthie, Susan J, Yamani, Naouale El, Engelward, Bevin, Gaivão, Isabel, Giovannelli, Lisa, Godschalk, Roger, Guilherme, Sofia, Gutzkow, Kristine B, Habas, Khaled, Hernández, Alba, Herrero, Oscar, Isidori, Marina, Jha, Awadhesh N, Knasmüller, Siegfried, Kooter, Ingeborg M, Koppen, Gudrun, Kruszewski, Marcin, Ladeira, Carina, Laffon, Blanca, Larramendy, Marcelo, Hégarat, Ludovic Le, Lewies, Angélique, Lewinska, Anna, Liwszyc, Guillermo E, de Cerain, Adela López, Manjanatha, Mugimane, Marcos, Ricard, Milić, Mirta, de Andrade, Vanessa Morae, Moretti, Massimo, Muruzabal, Damian, Novak, Matjaž, Oliveira, Rui, Olsen, Ann-Karin, Owiti, Norah, Pacheco, Mário, Pandey, Alok K, Pfuhler, Stefan, Pourrut, Bertrand, Reisinger, Kerstin, Rojas, Emilio, Rundén-Pran, Elise, Sanz-Serrano, Julen, Shaposhnikov, Sergey, Sipinen, Ville, Smeets, Karen, Stopper, Helga, Teixeira, João Paulo, Valdiglesias, Vanessa, Valverde, Mahara, van Acker, Frederique, van Schooten, Frederik-Jan, Vasquez, Marie, Wentzel, Johannes F, Wnuk, Maciej, Wouters, Annelie, Žegura, Bojana, Zikmund, Toma, Langie, Sabine A S, and Azqueta, Amaya
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Comet assay, DNA damage, animals, plants ,ENABLES HIGH-THROUGHPUT ,[SDV]Life Sciences [q-bio] ,INDUCED OXIDATIVE STRESS ,alkaline comet assay ,protocols ,EPITHELIAL-CELLS ,DOUBLE-STRAND BREAKS ,Article ,General Biochemistry, Genetics and Molecular Biology ,INTER-LABORATORY VARIATION ,DROSOPHILA-MELANOGASTER ,CELL GEL-ELECTROPHORESIS ,IN-VIVO MODEL ,CORD BLOOD ,INTERSTRAND CROSS-LINKS - 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.The comet assay is commonly used to assess DNA damage. This collection of consensus protocols includes adaptations for a wide range of species and sample types, assay formats and detection of different types of DNA lesions.
- Published
- 2023