title: Studying the organization of DNA repair by single-cell and single-molecule imaging.
abstract: DNA repair safeguards the genome against a diversity of DNA damaging agents. Although the mechanisms of many repair proteins have been examined separately in vitro, far less is known about the coordinated function of the whole repair machinery in vivo. Furthermore, single-cell studies indicate that DNA damage responses generate substantial variation in repair activities across cells. This review focuses on fluorescence imaging methods that offer a quantitative description of DNA repair in single cells by measuring protein concentrations, diffusion characteristics, localizations, interactions, and enzymatic rates. Emerging single-molecule and super-resolution microscopy methods now permit direct visualization of individual proteins and DNA repair events in vivo. We expect much can be learned about the organization of DNA repair by linking cell heterogeneity to mechanistic observations at the molecular level.
authors.affiliation: Department of Biochemistry, University of Oxford, Oxford OX1 3QU, United Kingdom. Electronic address: stephan.uphoff@bioch.ox.ac.uk.
authors.forename: Stephan
authors.initials: S
authors.lastname: Uphoff
authors.affiliation: Biological Physics Research Group, Clarendon Laboratory, Department of Physics, University of Oxford, Oxford OX1 3PU, United Kingdom. Electronic address: a.kapanidis1@physics.ox.ac.uk.
authors.forename: Achillefs N
authors.initials: AN
authors.lastname: Kapanidis
mesh_terms: D001419:Bacteria / Q000737:chemistry / Q000201:enzymology / Q000235:genetics; D004260:DNA Repair; D045643:DNA Repair Enzymes / Q000378:metabolism; D036681:Fluorescence Recovery After Photobleaching / Q000379:methods*; D008856:Microscopy, Fluorescence / Q000379:methods; D059010:Single-Cell Analysis / Q000379:methods*
chemical_list: D045643:DNA Repair Enzymes
keywords: Cell heterogeneity; DNA damage responses; DNA repair; Single-cell imaging; Single-molecule fluorescence; Super-resolution microscopy