title: Genome-wide mapping of DNA strand breaks.
abstract: Determination of cellular DNA damage has so far been limited to global assessment of genome integrity whereas nucleotide-level mapping has been restricted to specific loci by the use of specific primers. Therefore, only limited DNA sequences can be studied and novel regions of genomic instability can hardly be discovered. Using a well-characterized yeast model, we describe a straightforward strategy to map genome-wide DNA strand breaks without compromising nucleotide-level resolution. This technique, termed "damaged DNA immunoprecipitation" (dDIP), uses immunoprecipitation and the terminal deoxynucleotidyl transferase-mediated dUTP-biotin end-labeling (TUNEL) to capture DNA at break sites. When used in combination with microarray or next-generation sequencing technologies, dDIP will allow researchers to map genome-wide DNA strand breaks as well as other types of DNA damage and to establish a clear profiling of altered genes and/or intergenic sequences in various experimental conditions. This mapping technique could find several applications for instance in the study of aging, genotoxic drug screening, cancer, meiosis, radiation and oxidative DNA damage.
authors.affiliation: Département de Biochimie, Faculté de Médecine et des Sciences de la Santé, Université de Sherbrooke, Sherbrooke, Québec, Canada.
authors.forename: Frédéric
authors.initials: F
authors.lastname: Leduc
authors.forename: David
authors.initials: D
authors.lastname: Faucher
authors.forename: Geneviève
authors.initials: G
authors.lastname: Bikond Nkoma
authors.forename: Marie-Chantal
authors.initials: MC
authors.lastname: Grégoire
authors.forename: Mélina
authors.initials: M
authors.lastname: Arguin
authors.forename: Raymund J
authors.initials: RJ
authors.lastname: Wellinger
authors.forename: Guylain
authors.initials: G
authors.lastname: Boissonneault
mesh_terms: D002874:Chromosome Mapping / Q000379:methods*; D015825:Chromosomes, Fungal / Q000235:genetics / Q000378:metabolism; D053960:DNA Breaks; D004249:DNA Damage / Q000235:genetics / Q000502:physiology; D049770:Genes, Mating Type, Fungal / Q000235:genetics; D056426:Genetic Loci; D020287:In Situ Nick-End Labeling; D008954:Models, Biological; D030781:Organisms, Genetically Modified; D010957:Plasmids / Q000235:genetics; D016133:Polymerase Chain Reaction; D012441:Saccharomyces cerevisiae / Q000235:genetics*; D017422:Sequence Analysis, DNA