title: Measurement of DNA interstrand crosslinking in individual cells using the Single Cell Gel Electrophoresis (Comet) assay.
abstract: The Single Cell Gel Electrophoresis (Comet) assay, originally developed to allow visualisation of DNA strand break damage in individual cells, has been adapted to measure DNA interstrand cross-links. DNA interstrand cross-links are formed in cells by a number of commonly used cancer chemotherapy agents and are considered to be the critical lesion formed by such agents. This technique allows the analysis of DNA interstrand cross-link formation and repair at a single cell level, requires few cells, allows the determination of heterogeneity of response within a cell population and is sensitive enough to measure DNA interstrand cross-links at pharmacologically relevant doses. The method can be applied to any in vitro or in vivo application where a single cell suspension can be obtained. The method has also become invaluable in studies using human tissue and can be used as a method for pharmacodynamic analysis in early clinical trials.
authors.affiliation: Cancer Research UK Drug-DNA Interactions Research Group, UCL Cancer Institute, University College London, Paul O'Gorman Building, London, UK.
authors.forename: Victoria J
authors.initials: VJ
authors.lastname: Spanswick
authors.forename: Janet M
authors.initials: JM
authors.lastname: Hartley
authors.forename: John A
authors.initials: JA
authors.lastname: Hartley
mesh_terms: D001201:Ascites / Q000378:metabolism; D045744:Cell Line, Tumor; D020552:Comet Assay / Q000379:methods*; D003432:Cross-Linking Reagents / Q000494:pharmacology*; D004247:DNA / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D004249:DNA Damage; D006801:Humans; D008214:Lymphocytes / Q000187:drug effects / Q000378:metabolism; D008558:Melphalan / Q000494:pharmacology; D013194:Staining and Labeling
chemical_list: D003432:Cross-Linking Reagents; D004247:DNA; D008558:Melphalan