Double-strand breaks (DSBs) and their repair mechanisms are essential for normal cell life. However, quantitative analysis of DSBs on mammalian whole chromosomes remains difficult. The method described here enables the quantitative detection of mammalian chromosomal DSBs by pulsed-field gel electrophoresis (PFGE) using a contour-clamped homogeneous electric field (CHEF). We illustrate this method by measuring DNA damage-induced DSBs in mammalian cells. The electrophoresis conditions presented here enabled the visualization of fragmented DNA (several mega-base pairs down to 500 kbp) as a single band. Using this protocol, about 10-45 samples can be analyzed on a single gel, depending on the direction of electrophoresis.
Download full-text PDF |
Source |
|---|---|
| http://dx.doi.org/10.1007/978-1-0716-0323-9_9 | DOI Listing |
Publication Analysis
Top Keywords
Similar Publications
DNA damage and its links to neuronal aging and degeneration.
Neuron
January 2025
Departments of Psychiatry, Neuroscience, and Cell Biology, University of Texas Southwestern Medical Center, Dallas, TX, USA; Peter O' Donnell Brain Institute, University of Texas Southwestern Medical Center, Dallas, TX, USA. Electronic address:
DNA damage is a major risk factor for the decline of neuronal functions with age and in neurodegenerative diseases. While how DNA damage causes neurodegeneration is still being investigated, innovations over the past decade have provided significant insights into this issue. Breakthroughs in next-generation sequencing methods have begun to reveal the characteristics of neuronal DNA damage hotspots and the causes of DNA damage.
View Article and Find Full Text PDFHMGA1 Plays a Role in Counteracting DNA Damage Induced by BoHV-1 Productive Infection.
Int J Mol Sci
December 2024
College of Life Sciences, Hebei University, Baoding 071002, China.
Bovine herpesvirus 1 (BoHV-1) productive infection induces the generation of DNA double-strand breaks (DSBs), which may consequently lead to cell apoptosis. In response to DSBs, the DNA damage repair-related protein 53BP1 is recruited to the sites of DSBs, leading to the formation of 53BP1foci, which are crucial for the repair of damaged DNA and maintaining genomic integrity by repairing DSBs. In this study, we discovered that HMGA1 may play a significant role in counteracting virus infection-induced DNA damage, as the siRNA-mediated knockdown of HMGA1 protein expression or inhibition of HMGA1 activity by the chemical inhibitor Netropsin uniformly exacerbates the DNA damage induced by BoHV-1 productive infection.
View Article and Find Full Text PDFEfficacy of natural NF-κB inhibitors in the treatment of fibrosarcoma: an in vitro model study.
Front Cell Dev Biol
December 2024
Department of Molecular and Cellular Biology, Faculty of Pharmacy, Wroclaw Medical University, Wroclaw, Poland.
Introduction: NF-κB plays a pivotal role in the progression of cancers, including myosarcomas such as fibrosarcoma. Plants possess considerable potential for the provision of chemotherapeutic effects against cancer. The present study assessed, among others, the cytotoxicity, migration capacity and DNA damage induced by several natural compounds (berberine, curcumin, biochanin A, cucurbitacin E (CurE) and phenethyl caffeic acid (CAPE)) in cancer cells (WEHI-164) and normal muscle cells (L6).
View Article and Find Full Text PDFExpression Analysis of Cyst Specific Protein (CSP21) and Cellulose Synthase II (CSII) Genes in Acanthamoeba castellanii Trophozoites Exposed to Silver Nanoparticles Conjugated with Elaeagnus umbellata.
Acta Parasitol
January 2025
Department of Molecular Biology and Genetics, Ordu University, Ordu, Turkey.
Article Synopsis
- Acanthamoeba species are protozoa that can cause serious eye and CNS infections, and current treatments are often ineffective, especially in specific areas like the eye.
- The study evaluates the effectiveness of ethanolic fruit extract of E. umbellata, silver nanoparticles derived from it, and lauric acid in killing Acanthamoeba trophozoites and protecting DNA from damage.
- Results show that these treatments can significantly kill trophozoites and prevent DNA damage, suggesting potential new options for treating Acanthamoeba infections.
Wavelength-dependent DNA damage induced by single wavelengths of UV-C radiation (215 to 255 nm) in a human cornea model.
Sci Rep
January 2025
Center for Radiological Research, Columbia University Irving Medical Center, 630 West 168th Street, New York, 10032, USA.
Scientific bodies overseeing UV radiation protection recommend safety limits for exposure to ultraviolet radiation in the workplace based on published peer-reviewed data. To support this goal, a 3D model of the human cornea was used to assess the wavelength dependence of corneal damage induced by UV-C radiation. In the first set of experiments the models were exposed with or without simulated tears; at each wavelength (215-255 nm) cells with DNA dimers and their distribution within the epithelium were measured.
View Article and Find Full Text PDFWant AI Summaries of new PubMed Abstracts delivered to your In-box?
Enter search terms and have AI summaries delivered each week - change queries or unsubscribe any time!