TY - JOUR
T1 - Measuring DNA modifications with the comet assay
T2 - a compendium of protocols
AU - Collins, Andrew
AU - Møller, Peter
AU - Gajski, Goran
AU - Vodenková, Soňa
AU - Abdulwahed, Abdulhadi
AU - Anderson, Diana
AU - Bankoglu, Ezgi Eyluel
AU - Bonassi, Stefano
AU - Boutet-Robinet, Elisa
AU - Brunborg, Gunnar
AU - Chao, Christy
AU - Cooke, Marcus S
AU - Costa, Carla
AU - Costa, Solange
AU - Dhawan, Alok
AU - de Lapuente, Joaquin
AU - Bo', Cristian Del
AU - Dubus, Julien
AU - Dusinska, Maria
AU - Duthie, Susan J
AU - Yamani, Naouale El
AU - Engelward, Bevin
AU - Gaivão, Isabel
AU - Giovannelli, Lisa
AU - Godschalk, Roger
AU - Guilherme, Sofia
AU - Gutzkow, Kristine B
AU - Habas, Khaled
AU - Hernández, Alba
AU - Herrero, Oscar
AU - Isidori, Marina
AU - Jha, Awadhesh N
AU - Knasmüller, Siegfried
AU - Kooter, Ingeborg M
AU - Koppen, Gudrun
AU - Kruszewski, Marcin
AU - Ladeira, Carina
AU - Laffon, Blanca
AU - Larramendy, Marcelo
AU - Hégarat, Ludovic Le
AU - Lewies, Angélique
AU - Lewinska, Anna
AU - Liwszyc, Guillermo E
AU - de Cerain, Adela López
AU - Manjanatha, Mugimane
AU - Marcos, Ricard
AU - Milić, Mirta
AU - de Andrade, Vanessa Moraes
AU - Moretti, Massimo
AU - Muruzabal, Damian
AU - Marcos Dauder, Ricardo
N1 - Funding Information: 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. This paper reflects the views of the authors and does not necessarily reflect those of the US Food and Drug Administration or the National Institutes of Health. Publisher Copyright: © 2023, Springer Nature Limited.
PY - 2023/1/27
Y1 - 2023/1/27
N2 - 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.
AB - 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.
KW - Animals
KW - Comet Assay/methods
KW - DNA Damage
KW - DNA/genetics
KW - Eukaryotic Cells
KW - Humans
KW - Pyrimidine Dimers
UR - http://www.scopus.com/inward/record.url?scp=85146929563&partnerID=8YFLogxK
UR - https://www.mendeley.com/catalogue/f38ab80a-f8f9-3c7f-a3fb-56e0c82d1611/
U2 - 10.1038/s41596-022-00754-y
DO - 10.1038/s41596-022-00754-y
M3 - Review article
C2 - 36707722
AN - SCOPUS:85146929563
SN - 1754-2189
VL - 18
SP - 929
EP - 989
JO - Nature Protocols
JF - Nature Protocols
IS - 3
ER -