Measuring DNA modifications with the comet assay: a compendium of protocols
Autor(a) principal: | |
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Data de Publicação: | 2023 |
Outros Autores: | , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , |
Tipo de documento: | Artigo |
Idioma: | eng |
Título da fonte: | Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos) |
Texto Completo: | http://hdl.handle.net/10400.18/9048 |
Resumo: | 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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Measuring DNA modifications with the comet assay: a compendium of protocolsDNA DamageComet AssayEukaryotic CellsGeneticsGenotoxicidade AmbientalThe 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.We thank the hCOMET project (COST Action, CA 15132) for support. A. Azqueta thanks the Ministry of Science and Innovation (AGL2015-70640-R and PID2020-115348RB-I00) of the Spanish Government. S.G. thanks the national funds (OE), through FCT— Fundação para a Ciência e a Tecnologia (IP, in the scope of the framework contract foreseen in the numbers 4, 5 and 6 of the article 23, of the Decree-Law 57/2016, of 29 August, changed by Law 57/2017, of 19 July) for personal support. V.M.d.A. thanks the National Council of Technological and Scientific Development (CNPq—304203/2018-1) for personal support. D.M. thanks the program ‘Ayudas para la formación de profesorado universitario (FPU)’ of the Spanish Government for the predoctoral grant received. N.O. thanks the NIEHS Superfund Research Program ES ES027707 for personal support. J.S.-S. thanks the Government of Navarra for the predoctoral grant received. V.V. thanks the Ministerio de Educación, Cultura y Deporte (‘Beatriz Galindo’ program, BEAGAL18/00142) of the Spanish Government for personal support. M.S.C. acknowledges personal support from the National Institute of Environmental Health Sciences of the National Institutes of Health under award number: 1R41ES030274-01.Nature ResearchRepositório Científico do Instituto Nacional de SaúdeCollins, AndrewMøller, PeterGajski, GoranVodenková, SoňaAbdulwahed, AbdulhadiAnderson, DianaBankoglu, Ezgi EyluelBonassi, StefanoBoutet-Robinet, ElisaBrunborg, GunnarChao, ChristyCooke, Marcus S.Costa, CarlaCosta, SolangeDhawan, Alokde Lapuente, JoaquinBo’, Cristian DelDubus, JulienDusinska, MariaDuthie, Susan J.Yamani, Naouale ElEngelward, BevinGaivão, IsabelGiovannelli, LisaGodschalk, RogerGuilherme, SofiaGutzkow, Kristine B.Habas, KhaledHernández, AlbaHerrero, OscarIsidori, MarinaJha, Awadhesh N.Knasmüller, SiegfriedKooter, Ingeborg M.Koppen, GudrunKruszewski, MarcinLadeira, CarinaLaffon, BlancaLarramendy, MarceloHégarat, Ludovic LeLewies, AngéliqueLewinska, AnnaLiwszyc, Guillermo E.de Cerain, Adela LópezManjanatha, MugimaneMarcos, RicardMilić, Mirtade Andrade, Vanessa MoraesMoretti, MassimoMuruzabal, DamianNovak, MatjažOliveira, RuiOlsen, Ann-KarinOwiti, NorahPacheco, MárioPandey, Alok K.Pfuhler, StefanPourrut, BertrandReisinger, KerstinRojas, EmilioRundén-Pran, EliseSanz-Serrano, JulenShaposhnikov, SergeySipinen, VilleSmeets, KarenStopper, HelgaTeixeira, João PauloValdiglesias, VanessaValverde, Maharavan Acker, Frederiquevan Schooten, Frederik-JanVasquez, MarieWentzel, Johannes F.Wnuk, MaciejWouters, AnneliesŽegura, BojanaZikmund, TomasLangie, Sabine A.S.Azqueta, Amaya2024-02-06T16:07:52Z2023-03-272023-03-27T00:00:00Zinfo:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/articleapplication/pdfhttp://hdl.handle.net/10400.18/9048engNat Protoc. 2023 Mar;18(3):929-989. doi: 10.1038/s41596-022-00754-y. Epub 2023 Jan 27. Review1754-218910.1038/s41596-022-00754-ymetadata only accessinfo:eu-repo/semantics/openAccessreponame:Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos)instname:Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informaçãoinstacron:RCAAP2024-02-10T01:30:47Zoai:repositorio.insa.pt:10400.18/9048Portal AgregadorONGhttps://www.rcaap.pt/oai/openaireopendoar:71602024-03-20T02:37:21.795606Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos) - Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informaçãofalse |
dc.title.none.fl_str_mv |
Measuring DNA modifications with the comet assay: a compendium of protocols |
title |
Measuring DNA modifications with the comet assay: a compendium of protocols |
spellingShingle |
Measuring DNA modifications with the comet assay: a compendium of protocols Collins, Andrew DNA Damage Comet Assay Eukaryotic Cells Genetics Genotoxicidade Ambiental |
title_short |
Measuring DNA modifications with the comet assay: a compendium of protocols |
title_full |
Measuring DNA modifications with the comet assay: a compendium of protocols |
title_fullStr |
Measuring DNA modifications with the comet assay: a compendium of protocols |
title_full_unstemmed |
Measuring DNA modifications with the comet assay: a compendium of protocols |
title_sort |
Measuring DNA modifications with the comet assay: a compendium of protocols |
author |
Collins, Andrew |
author_facet |
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 Moraes 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, Annelies Žegura, Bojana Zikmund, Tomas Langie, Sabine A.S. Azqueta, Amaya |
author_role |
author |
author2 |
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 Moraes 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, Annelies Žegura, Bojana Zikmund, Tomas Langie, Sabine A.S. Azqueta, Amaya |
author2_role |
author author author author author author author author author author author author author author author author author author author author author author author author author author author author author author author author author author author author author author author author author author author author author author author author author author author author author author author author author author author author author author author author author author author author author author author author author author author author author author |
dc.contributor.none.fl_str_mv |
Repositório Científico do Instituto Nacional de Saúde |
dc.contributor.author.fl_str_mv |
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 Moraes 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, Annelies Žegura, Bojana Zikmund, Tomas Langie, Sabine A.S. Azqueta, Amaya |
dc.subject.por.fl_str_mv |
DNA Damage Comet Assay Eukaryotic Cells Genetics Genotoxicidade Ambiental |
topic |
DNA Damage Comet Assay Eukaryotic Cells Genetics Genotoxicidade Ambiental |
description |
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. |
publishDate |
2023 |
dc.date.none.fl_str_mv |
2023-03-27 2023-03-27T00:00:00Z 2024-02-06T16:07:52Z |
dc.type.status.fl_str_mv |
info:eu-repo/semantics/publishedVersion |
dc.type.driver.fl_str_mv |
info:eu-repo/semantics/article |
format |
article |
status_str |
publishedVersion |
dc.identifier.uri.fl_str_mv |
http://hdl.handle.net/10400.18/9048 |
url |
http://hdl.handle.net/10400.18/9048 |
dc.language.iso.fl_str_mv |
eng |
language |
eng |
dc.relation.none.fl_str_mv |
Nat Protoc. 2023 Mar;18(3):929-989. doi: 10.1038/s41596-022-00754-y. Epub 2023 Jan 27. Review 1754-2189 10.1038/s41596-022-00754-y |
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metadata only access info:eu-repo/semantics/openAccess |
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Nature Research |
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Nature Research |
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