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Single-Molecule Dynamics and Localization of DNA Repair Proteins in Cells.
- Source :
-
Methods in enzymology [Methods Enzymol] 2018; Vol. 600, pp. 375-406. Date of Electronic Publication: 2018 Feb 01. - Publication Year :
- 2018
-
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.<br /> (© 2018 Elsevier Inc. All rights reserved.)
- 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
Subjects
Details
- Language :
- English
- ISSN :
- 1557-7988
- Volume :
- 600
- Database :
- MEDLINE
- Journal :
- Methods in enzymology
- Publication Type :
- Academic Journal
- Accession number :
- 29458767
- Full Text :
- https://doi.org/10.1016/bs.mie.2017.11.015