Measuring DNA modifications with the comet assay: a compendium of protocols - INRAE - Institut national de recherche pour l’agriculture, l’alimentation et l’environnement
Article Dans Une Revue Nature Protocols Année : 2023

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

Andrew Collins (1) , Peter Møller (2) , Goran Gajski (3) , Soňa Vodenková (4, 5) , Abdulhadi Abdulwahed (6) , Diana Anderson (7) , Ezgi Eyluel Bankoglu (8) , Stefano Bonassi (9, 10) , Elisa Boutet-Robinet (11) , Gunnar Brunborg (12, 13) , Christy Chao (14) , Marcus Cooke (15) , Carla Costa (16, 17) , Solange Costa (16, 17) , Alok Dhawan (18) , Joaquin de Lapuente (19) , Cristian Del Bo’ (20) , Julien Dubus (21, 22) , Maria Dusinska (23, 24) , Susan Duthie (25) , Naouale El Yamani (24) , Bevin Engelward (26) , Isabel Gaivão (27) , Lisa Giovannelli (28) , Roger Godschalk (29) , Sofia Guilherme (30) , Kristine Gutzkow (31, 12) , Khaled Habas (32, 7) , Alba Hernández (33, 34) , Oscar Herrero (35) , Marina Isidori (36) , Awadhesh Jha (37, 38) , Siegfried Knasmüller (39) , Ingeborg Kooter (40) , Gudrun Koppen (41) , Marcin Kruszewski (42, 43) , Carina Ladeira (44) , Blanca Laffon , Marcelo Larramendy , Ludovic Le Hégarat (45) , Angélique Lewies , Anna Lewinska , Guillermo 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 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 Wentzel , Maciej Wnuk , Annelies Wouters , Bojana Žegura , Tomas Zikmund , Sabine Langie , Amaya Azqueta
1 UiO - University of Oslo
2 UCPH - University of Copenhagen = Københavns Universitet
3 Institute for Medical Research and Occupational Health
4 Institute of Experimental Medicine of the Academy of Sciences of the Czech Republic
5 UK - Univerzita Karlova [Praha, Česká republika] = Charles University [Prague, Czech Republic]
6 FIU - Florida International University [Miami]
7 University of Bradford
8 JMU - Julius-Maximilians-Universität Würzburg = University of Würzburg
9 IRCCS San Raffaele Pisana - Istituto di Ricovero e Cura a Carattere Scientifico
10 UniSR - Universita Vita Salute San Raffaele = Vita-Salute San Raffaele University [Milan, Italie]
11 ToxAlim-COMICS - Contaminants & Stress Cellulaire
12 Centre for Environmental Radioactivity (CERAD CoE)
13 Norvegian Institut of Public Health
14 MIT - Massachusetts Institute of Technology
15 USF - University of South Florida [Tampa]
16 INSA - Instituto Nacional de Saùde Dr Ricardo Jorge [Portugal]
17 Universidade do Porto = University of Porto
18 Centre for Biomedical Research, University of Hull,
19 AC Marca
20 UNIMI - Università degli Studi di Milano = University of Milan
21 BIAM - Institut de Biosciences et Biotechnologies d'Aix-Marseille (ex-IBEB)
22 SECR - Service d'Etudes du Comportement des Radionucléides
23 SCCS - Scientific Committee for Consumer Safety (SCCS)
24 NILU - Norwegian Institute for Air Research
25 University of Aberdeen
26 BE - Department of Biological Engineering [Cambridge]
27 UTAD - Universidade de Trás-os-Montes e Alto Douro
28 NEUROFARBA Department [Firenze, Italy]
29 NUTRIM - School of Nutrition and Translational Research in Metabolism [Maastricht]
30 CESAM - Centre for Environmental and Marine Studies [Aveiro]
31 NIPH - Norwegian Institute of Public Health [Oslo]
32 School of Chemistry and Biosciences
33 Department of Genetics, Microbiology and Statistics, and IRBio, University of Barcelona, Barcelona, Spain
34 University of Barcelona
35 UNED - Universidad Nacional de Educación a Distancia
36 Università degli studi della Campania "Luigi Vanvitelli" = University of the Study of Campania Luigi Vanvitelli
37 SBMS - School of Biological and Marine Science
38 Plymouth University
39 Medizinische Universität Wien = Medical University of Vienna
40 TNO - The Netherlands Organisation for Applied Scientific Research
41 VITO - Flemish Institute for Technological Research
42 INCT - Institute of Nuclear Chemistry and Technology
43 Institute of Rural Health
44 Escola Superior de Tecnologia da Saude de Lisboa
45 Laboratoire de Fougères - ANSES
Marcus Cooke
Joaquin de Lapuente
Susan Duthie
Blanca Laffon
Marcelo Larramendy
  • Fonction : Auteur
Ludovic Le Hégarat
Angélique Lewies
Anna Lewinska
  • Fonction : Auteur
Guillermo Liwszyc
  • Fonction : Auteur
Adela López de Cerain
  • Fonction : Auteur
Mugimane Manjanatha
  • Fonction : Auteur
Ricard Marcos
Mirta Milić
Vanessa Moraes de Andrade
Massimo Moretti
Damian Muruzabal
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Matjaž Novak
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Rui Oliveira
Ann-Karin Olsen
Norah Owiti
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Mário Pacheco
  • Fonction : Auteur
Alok Pandey
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Stefan Pfuhler
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Bertrand Pourrut
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Kerstin Reisinger
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Emilio Rojas
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Elise Rundén-Pran
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Julen Sanz-Serrano
Sergey Shaposhnikov
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Ville Sipinen
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Karen Smeets
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Helga Stopper
João Paulo Teixeira
Vanessa Valdiglesias
Mahara Valverde
Frederique van Acker
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Frederik-Jan van Schooten
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Marie Vasquez
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Johannes Wentzel
Maciej Wnuk
Annelies Wouters
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Bojana Žegura
Tomas Zikmund
Sabine Langie
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Amaya Azqueta
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Résumé

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.
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Dates et versions

hal-03997215 , version 1 (28-09-2023)

Identifiants

Citer

Andrew Collins, Peter Møller, Goran Gajski, Soňa Vodenková, Abdulhadi Abdulwahed, et al.. Measuring DNA modifications with the comet assay: a compendium of protocols. Nature Protocols, 2023, ⟨10.1038/s41596-022-00754-y⟩. ⟨hal-03997215⟩
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