A high-throughput 384-well CometChip platform reveals a role for 3-methyladenine in the cellular response to etoposide-induced DNA damage.
Journal
NAR genomics and bioinformatics
ISSN: 2631-9268
Titre abrégé: NAR Genom Bioinform
Pays: England
ID NLM: 101756213
Informations de publication
Date de publication:
Sep 2022
Sep 2022
Historique:
received:
03
04
2022
revised:
11
08
2022
accepted:
05
09
2022
entrez:
16
9
2022
pubmed:
17
9
2022
medline:
17
9
2022
Statut:
epublish
Résumé
The Comet or single-cell gel electrophoresis assay is a highly sensitive method to measure cellular, nuclear genome damage. However, low throughput can limit its application for large-scale studies. To overcome these limitations, a 96-well CometChip platform was recently developed that increases throughput and reduces variation due to simultaneous processing and automated analysis of 96 samples. To advance throughput further, we developed a 384-well CometChip platform that allows analysis of ∼100 cells per well. The 384-well CometChip extends the capacity by 4-fold as compared to the 96-well system, enhancing application for larger DNA damage analysis studies. The overall sensitivity of the 384-well CometChip is consistent with that of the 96-well system, sensitive to genotoxin exposure and to loss of DNA repair capacity. We then applied the 384-well platform to screen a library of protein kinase inhibitors to probe each as enhancers of etoposide induced DNA damage. Here, we found that 3-methyladenine significantly increased levels of etoposide-induced DNA damage. Our results suggest that a 384-well CometChip is useful for large-scale DNA damage analyses, which may have increased potential in the evaluation of chemotherapy efficacy, compound library screens, population-based analyses of genome damage and evaluating the impact of environmental genotoxins on genome integrity.
Identifiants
pubmed: 36110898
doi: 10.1093/nargab/lqac065
pii: lqac065
pmc: PMC9469923
doi:
Types de publication
Journal Article
Langues
eng
Pagination
lqac065Subventions
Organisme : NCI NIH HHS
ID : R01 CA148629
Pays : United States
Organisme : NCI NIH HHS
ID : R01 CA238061
Pays : United States
Organisme : NIEHS NIH HHS
ID : R44 ES032522
Pays : United States
Organisme : NIEHS NIH HHS
ID : U01 ES029518
Pays : United States
Informations de copyright
© The Author(s) 2022. Published by Oxford University Press on behalf of NAR Genomics and Bioinformatics.
Références
J Biol Chem. 2010 Apr 2;285(14):10850-61
pubmed: 20123989
Environ Mol Mutagen. 2010 Jul;51(6):520-6
pubmed: 20658645
Front Genet. 2013 Feb 25;4:19
pubmed: 23443494
Free Radic Biol Med. 2021 Oct;174:89-99
pubmed: 34324980
Nucleic Acids Res. 2009 Dec;37(22):e150
pubmed: 19828597
Mol Biotechnol. 2004 Mar;26(3):249-61
pubmed: 15004294
Exp Cell Res. 1988 Mar;175(1):184-91
pubmed: 3345800
Biochem Biophys Res Commun. 1984 Aug 30;123(1):291-8
pubmed: 6477583
Mutat Res. 2001 Oct 18;497(1-2):169-75
pubmed: 11525920
Genes (Basel). 2019 Oct 30;10(11):
pubmed: 31671674
Biochemistry. 2005 Aug 9;44(31):10613-9
pubmed: 16060670
Proc Natl Acad Sci U S A. 2010 Jun 1;107(22):10008-13
pubmed: 20534572
Cancer. 1991 Jan 1;67(1 Suppl):215-9
pubmed: 1984822
Environ Mol Mutagen. 2018 May;59(4):322-333
pubmed: 29536573
Mutagenesis. 2015 Jan;30(1):11-9
pubmed: 25527723
Front Pharmacol. 2013 Jan 31;4:5
pubmed: 23386830
J Chromatogr B Biomed Sci Appl. 1999 Feb 5;722(1-2):225-54
pubmed: 10068143
Cell Rep. 2021 Nov 2;37(5):109917
pubmed: 34731617
Mol Cancer Res. 2012 Jul;10(7):945-57
pubmed: 22596249
Clin Cancer Res. 2006 Jul 1;12(13):4119-26
pubmed: 16818713
Biochem Biophys Res Commun. 2004 Dec 24;325(4):1279-85
pubmed: 15555565
Nat Rev Cancer. 2012 Sep;12(9):587-98
pubmed: 22918414
J Biol Chem. 2007 Jul 20;282(29):21206-12
pubmed: 17513297
Nucleic Acids Res. 2020 Feb 20;48(3):e13
pubmed: 31822921
Best Pract Res Clin Endocrinol Metab. 2020 May;34(3):101434
pubmed: 32622829
Environ Mol Mutagen. 2017 Aug;58(7):508-521
pubmed: 28755435
Mutagenesis. 2011 Nov;26(6):689-95
pubmed: 21778357
Autophagy. 2017;13(11):1828-1840
pubmed: 28837411
Nature. 2009 Oct 22;461(7267):1071-8
pubmed: 19847258
Biochemistry (Mosc). 2007 Aug;72(8):878-86
pubmed: 17922646
Mol Cancer Ther. 2008 Jul;7(7):1772-81
pubmed: 18644989
Sci Rep. 2018 Feb 9;8(1):2771
pubmed: 29426857
DNA Repair (Amst). 2007 Jun 1;6(6):695-711
pubmed: 17337257
Nature. 2017 Jan 5;541(7635):87-91
pubmed: 28002403
Mutagenesis. 2012 Nov;27(6):665-72
pubmed: 22844078
J Cell Biochem. 2005 Jul 1;95(4):794-804
pubmed: 15838887
Methods Mol Biol. 2002;203:179-94
pubmed: 12073441
Cancer Radiother. 1999 Jul-Aug;3(4):289-95
pubmed: 10486539
Lancet Oncol. 2010 Feb;11(2):184-92
pubmed: 20152770
J Immunol. 2012 Oct 15;189(8):4154-64
pubmed: 22972931
Eur J Cancer. 1998 Sep;34(10):1514-21
pubmed: 9893622
Mamm Genome. 2018 Feb;29(1-2):182-189
pubmed: 29299621
Int J Mol Med. 2011 Apr;27(4):599-606
pubmed: 21274505
Environ Mol Mutagen. 2018 Jul;59(6):529-538
pubmed: 29761828
Cell. 2001 Jan 12;104(1):107-17
pubmed: 11163244
Cell Cycle. 2013 Mar 15;12(6):907-15
pubmed: 23422001
J Chromatogr A. 2009 Mar 6;1216(10):1917-29
pubmed: 19100556
Environ Mol Mutagen. 2000;35(3):206-21
pubmed: 10737956
J Vis Exp. 2017 Oct 11;(128):
pubmed: 29053680
Front Genet. 2014 Aug 25;5:288
pubmed: 25202323
DNA Repair (Amst). 2011 Dec 10;10(12):1282-93
pubmed: 22041025