Abstract
A fully optimized staining method for detecting sister chromatid exchanges in cultured cells is presented. The method gives reproducibly robust quantitative results. Sister chromatid exchange is a classic toxicology assay for genotoxicity and for detecting alterations to the biochemistry underlying cellular homologous recombination. Growth of cells in the presence of 5′-bromo-deoxyuridine for two rounds of DNA replication followed by collecting metaphase spreads on glass slides, treatment with the UV-sensitive dye Hoechst 33258, long-wave UV light exposure, and Giemsa staining gives a permanent record of the exchanges.
This is a preview of subscription content, log in via an institution to check access.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
References
Rothkamm K, Kruger I, Thompson LH, Lobrich M (2003) Pathways of DNA double-strand break repair during the mammalian cell cycle. Mol Cell Biol 23:5706–5715
Helleday T (2010) Homologous recombination in cancer development, treatment and development of drug resistance. Carcinogenesis 31:955–960
Reliene R, Bishop AJ, Schiestl RH (2007) Involvement of homologous recombination in carcinogenesis. Adv Genet 58:67–87
Kyle S, Thomas HD, Mitchell J, Curtin NJ (2008) Exploiting the Achilles heel of cancer: the therapeutic potential of poly(ADP-ribose) polymerase inhibitors in BRCA2-defective cancer. Br J Radiol 81(Spec. No. 1):S6–S11
Ashworth A (2008) A synthetic lethal therapeutic approach: poly(ADP) ribose polymerase inhibitors for the treatment of cancers deficient in DNA double-strand break repair. J Clin Oncol 26:3785–3790
Schultz N, Lopez E, Saleh-Gohari N, Helleday T (2003) Poly(ADP-ribose) polymerase (PARP-1) has a controlling role in homologous recombination. Nucleic Acids Res 31: 4959–4964
Wilson DM III, Thompson LH (2007) Molecular mechanisms of sister-chromatid exchange. Mutat Res 616:11–23
Perry P, Evans HJ (1975) Cytological detection of mutagen-carcinogen exposure by sister chromatid exchange. Nature 258:121–125
Thompson LH (2005) Unraveling the Fanconi anemia–DNA repair connection. Nat Genet 37:921–922
Killen MW, Stults DM, Adachi N, Hanakahi L, Pierce AJ (2009) Loss of Bloom syndrome protein destabilizes human gene cluster architecture. Hum Mol Genet 18:3417–3428
Wolff S, Afzal V (1996) Segregation of DNA polynucleotide strands into sister chromatids and the use of endoreduplicated cells to track sister chromatid exchanges induced by crosslinks, alkylations, or x-ray damage. Proc Natl Acad Sci U S A 93:5765–5769
Perry P, Wolff S (1974) New Giemsa method for the differential staining of sister chromatids. Nature 251:156–158
Yankiwski V, Noonan JP, Neff NF (2001) The C-terminal domain of the Bloom syndrome DNA helicase is essential for genomic stability. BMC Cell Biol 2:11
Pinkel D, Thompson LH, Gray JW, Vanderlaan M (1985) Measurement of sister chromatid exchanges at very low bromodeoxyuridine substitution levels using a monoclonal antibody in Chinese hamster ovary cells. Cancer Res 45: 5795–5798
Stoddart MJ (2011) Cell viability assays: an introduction. Methods Mol Biol 740:1–6
Author information
Authors and Affiliations
Corresponding author
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2014 Humana Press
About this protocol
Cite this protocol
Stults, D.M., Killen, M.W., Pierce, A.J. (2014). The Sister Chromatid Exchange (SCE) Assay. In: Keohavong, P., Grant, S. (eds) Molecular Toxicology Protocols. Methods in Molecular Biology, vol 1105. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-62703-739-6_32
Download citation
DOI: https://doi.org/10.1007/978-1-62703-739-6_32
Published:
Publisher Name: Humana Press, Totowa, NJ
Print ISBN: 978-1-62703-738-9
Online ISBN: 978-1-62703-739-6
eBook Packages: Springer Protocols