ArticleMethods in enzymology2018
Quantitative Methods to Investigate the 4D Dynamics of Heterochromatic Repair Sites in Drosophila Cells.
Article in Methods in enzymology, 2018. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 18 papers.
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Who cites it
18 citing papers in PubMed, 23 citations in OpenAlex.
- Genome scale CRISPR screens identify actin capping proteins as key modulators of therapeutic responses to radiation and immunotherapy.bioRxiv : the preprint server for biology · 2025Article
- Nuclear and genome dynamics underlying DNA double-strand break repair.Nature reviews. Molecular cell biology · 2025Review
- Off-pore Nup98 condensates mobilize heterochromatic breaks and exclude Rad51.Molecular cell · 2025Article
- "Off-pore" nucleoporins relocalize heterochromatic breaks through phase separation.bioRxiv : the preprint server for biology · 2024Article
- Replication forks associated with long nuclear actin filaments in mild stress conditions display increased dynamics.microPublication biology · 2024Article
- Nup153 is not required for anchoring heterochromatic DSBs to the nuclear periphery.microPublication biology · 2024Article
- Nuclear actin polymerization rapidly mediates replication fork remodeling upon stress by limiting PrimPol activity.Nature communications · 2023Article
- Review
- Multi-scale dynamics of heterochromatin repair.Current opinion in genetics & development · 2021Review
- Article
- Live Cell Imaging of Nuclear Actin Filaments and Heterochromatic Repair foci in Drosophila and Mouse Cells.Methods in molecular biology (Clifton, N.J.) · 2021Article
- Complex Chromatin Motions for DNA Repair.Frontiers in genetics · 2020Review
- Arp2/3 and Unc45 maintain heterochromatin stability inExperimental biology and medicine (Maywood, N.J.) · 2019Article
- Actin' between phase separated domains for heterochromatin repair.DNA repair · 2019Review
- Nuclear actin filaments in DNA repair dynamics.Nature cell biology · 2019Review
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- Review
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Corrections and comments
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Authors and funding
7 authors at 1 institution in 1 country.
Funding
Abstract
Heterochromatin is mostly composed of long stretches of repeated DNA sequences prone to ectopic recombination during double-strand break (DSB) repair. In Drosophila, "safe" homologous recombination (HR) repair of heterochromatic DSBs relies on a striking relocalization of repair sites to the nuclear periphery. Central to understanding heterochromatin repair is the ability to investigate the 4D dynamics (movement in space and time) of repair sites. A specific challenge of these studies is preventing phototoxicity and photobleaching effects while imaging the sample over long periods of time, and with sufficient time points and Z-stacks to track repair foci over time. Here we describe an optimized approach for high-resolution live imaging of heterochromatic DSBs in Drosophila cells, with a specific emphasis on the fluorescent markers and imaging setup used to capture the motion of repair foci over long-time periods. We detail approaches that minimize photobleaching and phototoxicity with a DeltaVision widefield deconvolution microscope, and image processing techniques for signal recovery postimaging using SoftWorX and Imaris software. We present a method to derive mean square displacement curves revealing some of the biophysical properties of the motion. Finally, we describe a method in R to identify tracts of directed motions (DMs) in mixed trajectories. These approaches enable a deeper understanding of the mechanisms of heterochromatin dynamics and genome stability in the three-dimensional context of the nucleus and have broad applicability in the field of nuclear dynamics.
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