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

2023-03-27Journal article Restricted access
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dc.contributor.authorCollins, Andrew
dc.contributor.authorMøller, Peter
dc.contributor.authorGajski, Goran
dc.contributor.authorVodenková, Soňa
dc.contributor.authorAbdulwahed, Abdulhadi
dc.contributor.authorAnderson, Diana
dc.contributor.authorBankoglu, Ezgi Eyluel
dc.contributor.authorBonassi, Stefano
dc.contributor.authorBoutet-Robinet, Elisa
dc.contributor.authorBrunborg, Gunnar
dc.contributor.authorChao, Christy
dc.contributor.authorCooke, Marcus S.
dc.contributor.authorCosta, Carla
dc.contributor.authorCosta, Solange
dc.contributor.authorDhawan, Alok
dc.contributor.authorde Lapuente, Joaquin
dc.contributor.authorBo’, Cristian Del
dc.contributor.authorDubus, Julien
dc.contributor.authorDusinska, Maria
dc.contributor.authorDuthie, Susan J.
dc.contributor.authorYamani, Naouale El
dc.contributor.authorEngelward, Bevin
dc.contributor.authorGaivão, Isabel
dc.contributor.authorGiovannelli, Lisa
dc.contributor.authorGodschalk, Roger
dc.contributor.authorGuilherme, Sofia
dc.contributor.authorGutzkow, Kristine B.
dc.contributor.authorHabas, Khaled
dc.contributor.authorHernández, Alba
dc.contributor.authorHerrero, Oscar
dc.contributor.authorIsidori, Marina
dc.contributor.authorJha, Awadhesh N.
dc.contributor.authorKnasmüller, Siegfried
dc.contributor.authorKooter, Ingeborg M.
dc.contributor.authorKoppen, Gudrun
dc.contributor.authorKruszewski, Marcin
dc.contributor.authorLadeira, Carina
dc.contributor.authorLaffon, Blanca
dc.contributor.authorLarramendy, Marcelo
dc.contributor.authorHégarat, Ludovic Le
dc.contributor.authorLewies, Angélique
dc.contributor.authorLewinska, Anna
dc.contributor.authorLiwszyc, Guillermo E.
dc.contributor.authorde Cerain, Adela López
dc.contributor.authorManjanatha, Mugimane
dc.contributor.authorMarcos, Ricard
dc.contributor.authorMilić, Mirta
dc.contributor.authorde Andrade, Vanessa Moraes
dc.contributor.authorMoretti, Massimo
dc.contributor.authorMuruzabal, Damian
dc.contributor.authorNovak, Matjaž
dc.contributor.authorOliveira, Rui
dc.contributor.authorOlsen, Ann-Karin
dc.contributor.authorOwiti, Norah
dc.contributor.authorPacheco, Mário
dc.contributor.authorPandey, Alok K.
dc.contributor.authorPfuhler, Stefan
dc.contributor.authorPourrut, Bertrand
dc.contributor.authorReisinger, Kerstin
dc.contributor.authorRojas, Emilio
dc.contributor.authorRundén-Pran, Elise
dc.contributor.authorSanz-Serrano, Julen
dc.contributor.authorShaposhnikov, Sergey
dc.contributor.authorSipinen, Ville
dc.contributor.authorSmeets, Karen
dc.contributor.authorStopper, Helga
dc.contributor.authorTeixeira, João Paulo
dc.contributor.authorValdiglesias, Vanessa
dc.contributor.authorValverde, Mahara
dc.contributor.authorvan Acker, Frederique
dc.contributor.authorvan Schooten, Frederik-Jan
dc.contributor.authorVasquez, Marie
dc.contributor.authorWentzel, Johannes F.
dc.contributor.authorWnuk, Maciej
dc.contributor.authorWouters, Annelies
dc.contributor.authorŽegura, Bojana
dc.contributor.authorZikmund, Tomas
dc.contributor.authorLangie, Sabine A.S.
dc.contributor.authorAzqueta, Amaya
dc.date.accessioned2024-02-06T16:07:52Z
dc.date.available2024-02-06T16:07:52Z
dc.date.issued2023-03-27
dc.description.abstractThe 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.pt_PT
dc.description.sponsorshipWe 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.pt_PT
dc.description.versioninfo:eu-repo/semantics/publishedVersionpt_PT
dc.identifier.citationNat Protoc. 2023 Mar;18(3):929-989. doi: 10.1038/s41596-022-00754-y. Epub 2023 Jan 27. Reviewpt_PT
dc.identifier.doi10.1038/s41596-022-00754-ypt_PT
dc.identifier.issn1754-2189
dc.identifier.urihttp://hdl.handle.net/10400.18/9048
dc.language.isoengpt_PT
dc.peerreviewedyespt_PT
dc.publisherNature Researchpt_PT
dc.relation.publisherversionhttps://www.nature.com/articles/s41596-022-00754-ypt_PT
dc.rights.urihttp://creativecommons.org/licenses/by/4.0/pt_PT
dc.subjectDNA Damagept_PT
dc.subjectComet Assaypt_PT
dc.subjectEukaryotic Cellspt_PT
dc.subjectGeneticspt_PT
dc.subjectGenotoxicidade Ambientalpt_PT
dc.titleMeasuring DNA modifications with the comet assay: a compendium of protocolspt_PT
dc.typejournal article
dspace.entity.typePublication
oaire.citation.endPage989pt_PT
oaire.citation.issue3pt_PT
oaire.citation.startPage929pt_PT
oaire.citation.titleNature Protocolspt_PT
oaire.citation.volume18pt_PT
rcaap.embargofctAcesso de acordo com a política editorial da revista.pt_PT
rcaap.rightsclosedAccesspt_PT
rcaap.typearticlept_PT

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