Mutagenesis, Vol. 16, No. 4, 297-301,
July 2001
© 2001 UK Environmental Mutagen Society/Oxford University Press
Inter-individual differences in repair of DNA base oxidation, measured in vitro with the comet assay
inská,1
t
tina,4
Rowett Research Institute, Greenburn Road, Bucksburn, Aberdeen, AB21 9SB, UK
There is a need for a reliable, robust and sensitive assay for DNA repair, suitable for use with human lymphocyte samples in molecular epidemiological investigations. The comet assay (single cell alkaline gel electrophoresis) has been modified to measure the ability of a simple subcellular extract of lymphocytes to carry out the initial step of repair, i.e. incision, on a DNA substrate carrying specific lesions—namely, oxidized bases introduced by visible light in the presence of photosensitizer. The cell extract is free of non-specific nuclease activity, incising DNA only if the DNA has been treated with photosensitizer and light. The activity varies between individuals, but consistency is seen between samples from each individual taken on occasions several months apart. The lack of activity of extract from Ogg1– mouse cells (deficient in the glycosylase that excises 8-oxoguanine) in this assay confirms that the activity measured is predominantly excision repair of oxidized bases. This new DNA repair assay is simple, rapid and requires only small quantities of lymphocyte extract (obtainable from 10 ml blood).
1 Permanent addresses: Institute of Preventive and Clinical Medicine, Limbová 14, 833 01 Bratislava,
2 Cancer Research Institute, Vlárska 7, 833 91 Bratislava, Slovakia,
3 Department of Experimental Medicine, Section of General Pathology, University of Pavia, Piazza Botta, 10, 27100 Pavia, Italy and
4 Purkyne Military Medical Academy, Trebeá 1575, 500 01 Hradec Králové, Czech Republic
5 To whom correspondence should be addressed.
CiteULike 
Connotea 
Del.icio.us What's this?
This article has been cited by other articles:
|
|
 |
|
  K. Vande Loock, I. Decordier, R. Ciardelli, D. Haumont, and M. Kirsch-Volders An aphidicolin-block nucleotide excision repair assay measuring DNA incision and repair capacity Mutagenesis, October 20, 2009; (2009) gep039v1. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 M. Herrera, G. Dominguez, J. M. Garcia, C. Pena, C. Jimenez, J. Silva, V. Garcia, I. Gomez, R. Diaz, P. Martin, et al. Differences in Repair of DNA Cross-links between Lymphocytes and Epithelial Tumor Cells from Colon Cancer Patients Measured In vitro with the Comet Assay Clin. Cancer Res., September 1, 2009; 15(17): 5466 - 5472. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
A. Azqueta, Y. Lorenzo, and A. R. Collins In vitro comet assay for DNA repair: a warning concerning application to cultured cells Mutagenesis, July 1, 2009; 24(4): 379 - 381. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 G. Gajski and V. Garaj-Vrhovac Radioprotective Effects of Honeybee Venom (Apis mellifera) Against 915-MHz Microwave Radiation-Induced DNA Damage in Wistar Rat Lymphocytes: In Vitro Study International Journal of Toxicology, March 1, 2009; 28(2): 88 - 98. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 Y. Lorenzo, A. Azqueta, L. Luna, F. Bonilla, G. Dominguez, and A. R. Collins The carotenoid {beta}-cryptoxanthin stimulates the repair of DNA oxidation damage in addition to acting as an antioxidant in human cells Carcinogenesis, February 1, 2009; 30(2): 308 - 314. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
M. Staruchova, A. R. Collins, K. Volkovova, C. Mislanova, Z. Kovacikova, J. Tulinska, A. Kocan, L. Staruch, L. Wsolova, and M. Dusinska Occupational exposure to mineral fibres. Biomarkers of oxidative damage and antioxidant defence and associations with DNA damage and repair Mutagenesis, July 1, 2008; 23(4): 249 - 260. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
A. R. Collins, A. A. Oscoz, G. Brunborg, I. Gaivao, L. Giovannelli, M. Kruszewski, C. C. Smith, and R. Stetina The comet assay: topical issues Mutagenesis, May 1, 2008; 23(3): 143 - 151. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
L. Forchhammer, E. V. Brauner, J. K. Folkmann, P. H. Danielsen, C. Nielsen, A. Jensen, S. Loft, G. Friis, and P. Moller Variation in assessment of oxidatively damaged DNA in mononuclear blood cells by the comet assay with visual scoring Mutagenesis, May 1, 2008; 23(3): 223 - 231. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
M. Dusinska and A. R. Collins The comet assay in human biomonitoring: gene-environment interactions Mutagenesis, May 1, 2008; 23(3): 191 - 205. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 E. Cordelli, A. M. Fresegna, A. D'Alessio, P. Eleuteri, M. Spano, F. Pacchierotti, and P. Villani ReProComet: A New In Vitro Method to Assess DNA Damage in Mammalian Sperm Toxicol. Sci., October 1, 2007; 99(2): 545 - 552. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 N. Gungor, R. W.L. Godschalk, D. M. Pachen, F. J. Van Schooten, and A. M. Knaapen Activated neutrophils inhibit nucleotide excision repair in human pulmonary epithelial cells: role of myeloperoxidase FASEB J, August 1, 2007; 21(10): 2359 - 2367. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
P. Vodicka, R. Stetina, V. Polakova, E. Tulupova, A. Naccarati, L. Vodickova, R. Kumar, M. Hanova, B. Pardini, J. Slyskova, et al. Association of DNA repair polymorphisms with DNA repair functional outcomes in healthy human subjects Carcinogenesis, March 1, 2007; 28(3): 657 - 664. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
M. Dusinska, Z. Dzupinkova, L. Wsolova, V. Harrington, and A. R. Collins Possible involvement of XPA in repair of oxidative DNA damage deduced from analysis of damage, repair and genotype in a human population study Mutagenesis, May 1, 2006; 21(3): 205 - 211. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
S. A.S. Langie, A. M. Knaapen, K. J.J. Brauers, D. van Berlo, F.-J. van Schooten, and R. W.L. Godschalk Development and validation of a modified comet assay to phenotypically assess nucleotide excision repair Mutagenesis, March 1, 2006; 21(2): 153 - 158. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 A. J. Lee, N. J. Hodges, and J. K. Chipman Interindividual Variability in Response to Sodium Dichromate-Induced Oxidative DNA Damage: Role of the Ser326Cys Polymorphism in the DNA-Repair Protein of 8-Oxo-7,8-Dihydro-2'-Deoxyguanosine DNA Glycosylase 1 Cancer Epidemiol. Biomarkers Prev., February 1, 2005; 14(2): 497 - 505. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
P. Vodicka, R. Kumar, R. Stetina, S. Sanyal, P. Soucek, V. Haufroid, M. Dusinska, M. Kuricova, M. Zamecnikova, L. Musak, et al. Genetic polymorphisms in DNA repair genes and possible links with DNA repair rates, chromosomal aberrations and single-strand breaks in DNA Carcinogenesis, May 1, 2004; 25(5): 757 - 763. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
N. Karunasinghe, J. Ryan, J. Tuckey, J. Masters, M. Jamieson, L. C. Clarke, J. R. Marshall, and L. R. Ferguson DNA Stability and Serum Selenium Levels in a High-Risk Group for Prostate Cancer Cancer Epidemiol. Biomarkers Prev., March 1, 2004; 13(3): 391 - 397. [Abstract] [Full Text]
|
|
|
|
|
|
R. Alleva, M. Tomasetti, M. D. Solenghi, F. Stagni, F. Gamberini, A. Bassi, P. M. Fornasari, G. Fanelli, and B. Borghi Lymphocyte DNA damage precedes DNA repair or cell death after orthopaedic surgery under general anaesthesia Mutagenesis, September 1, 2003; 18(5): 423 - 428. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 J. R. Sparrow, J. Zhou, and B. Cai DNA Is a Target of the Photodynamic Effects Elicited in A2E-Laden RPE by Blue-Light Illumination Invest. Ophthalmol. Vis. Sci., May 1, 2003; 44(5): 2245 - 2251. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
A. R. Collins, V. Harrington, J. Drew, and R. Melvin Nutritional modulation of DNA repair in a human intervention study Carcinogenesis, March 1, 2003; 24(3): 511 - 515. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
A. Collins and V. Harrington Repair of oxidative DNA damage: assessing its contribution to cancer prevention Mutagenesis, November 1, 2002; 17(6): 489 - 493. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
S.J. Duthie, L. Pirie, A.McE. Jenkinson, and S. Narayanan Cryopreserved versus freshly isolated lymphocytes in human biomonitoring: endogenous and induced DNA damage, antioxidant status and repair capability Mutagenesis, May 1, 2002; 17(3): 211 - 214. [Abstract] [Full Text] [PDF]
|
|