1. Single-Molecule Dynamics and Localization of DNA Repair Proteins in Cells.
- Author
-
Paul MW, Zelensky AN, Wyman C, and Kanaar R
- Subjects
- Animals, BRCA2 Protein chemistry, BRCA2 Protein metabolism, CRISPR-Cas Systems genetics, Cell Culture Techniques instrumentation, Cells, Cultured, DNA Breaks, Double-Stranded, Fluorescence Recovery After Photobleaching instrumentation, Fluorescence Recovery After Photobleaching methods, Gene Editing methods, Green Fluorescent Proteins chemistry, Intravital Microscopy instrumentation, Luminescent Agents chemistry, Mice, Microscopy, Fluorescence instrumentation, Microscopy, Fluorescence methods, Mouse Embryonic Stem Cells, Protein Binding, Rad51 Recombinase chemistry, Rad51 Recombinase metabolism, Single Molecule Imaging instrumentation, BRCA2 Protein analysis, Cell Culture Techniques methods, Intravital Microscopy methods, Rad51 Recombinase analysis, Recombinational DNA Repair, Single Molecule Imaging methods
- Abstract
Direct observation of individual protein molecules in their native environment, at nanometer resolution, in a living cell, in motion is not only fascinating but also uniquely informative. Several recent major technological advances in genomic engineering, protein and synthetic fluorophore development, and light microscopy have dramatically increased the accessibility of this approach. This chapter describes the procedures for modifying endogenous genomic loci to producing fluorescently tagged proteins, their high-resolution visualization, and analysis of their dynamics in mammalian cells, using DNA repair proteins BRCA2 and RAD51 as an example., (© 2018 Elsevier Inc. All rights reserved.)
- Published
- 2018
- Full Text
- View/download PDF