Generation of 3' single-stranded DNA (ssDNA) at the ends of a double-strand break (DSB) is essential to initiate repair by homology-directed mechanisms. Here we describe a Southern blot-based method to visualize the generation of ssDNA at the ends of site-specific DSBs generated in the Saccharomyces cerevisiae genome.
Download full-text PDF |
Source |
|---|---|
| http://dx.doi.org/10.1007/978-1-0716-0644-5_3 | DOI Listing |
Publication Analysis
Top Keywords
Similar Publications
CRISPR-Enabled Autonomous Transposable Element (CREATE) for RNA-based gene editing and delivery.
EMBO Rep
January 2025
Myeloid Therapeutics Inc., Cambridge, MA, 02139, USA.
To address a wide range of genetic diseases, genome editing tools that can achieve targeted delivery of large genes without causing double-strand breaks (DSBs) or requiring DNA templates are necessary. Here, we introduce CRISPR-Enabled Autonomous Transposable Element (CREATE), a genome editing system that combines the programmability and precision of CRISPR/Cas9 with the RNA-mediated gene insertion capabilities of the human LINE-1 (L1) element. CREATE employs a modified L1 mRNA to carry a payload gene, and a Cas9 nickase to facilitate targeted editing by L1-mediated reverse transcription and integration without relying on DSBs or DNA templates.
View Article and Find Full Text PDFE-cigarettes increase the risk of adenoma formation in murine colorectal cancer model.
Arch Toxicol
January 2025
Department of Medicine, University of California, San Diego, CA, 92093, USA.
E-cigarettes (E.cigs) cause inflammation and damage to human organs, including the lungs and heart. In the gut, E.
View Article and Find Full Text PDFBiological Amyloids Chemically Damage DNA.
ACS Chem Neurosci
January 2025
Department of Life Sciences, Chalmers University of Technology, 412 96 Gothenburg, Sweden.
Amyloid fibrils are protein polymers noncovalently assembled through β-strands arranged in a cross-β structure. Biological amyloids were considered chemically inert until we and others recently demonstrated their ability to catalyze chemical reactions in vitro. To further explore the functional repertoire of amyloids, we here probe if fibrils of α-synuclein (αS) display chemical reactivity toward DNA.
View Article and Find Full Text PDFInactivation of GSK3β by Ser phosphorylation prevents thymocyte necroptosis and impacts Tcr repertoire diversity.
Cell Death Differ
January 2025
Department of Immunology and Microbiology, University of Colorado, Anschutz Medical Campus, Aurora, CO, 80045, USA.
The assembly of Tcrb and Tcra genes require double negative (DN) thymocytes to undergo multiple rounds of programmed DNA double-strand breaks (DSBs), followed by their efficient repair. However, mechanisms governing cell cycle checkpoints and specific survival pathways during the repair process remain unclear. Here, we report high-resolution scRNA-seq analyses of individually sorted mouse DN3 and DN4 thymocytes, which reveals a G2M cell cycle checkpoint, in addition to the known G1 checkpoint, during Tcrb and Tcra recombination.
View Article and Find Full Text PDFRapid quantitative analysis of double-stranded plasmid DNA with capillary gel electrophoresis for applications in quality control and radiation research.
Sci Rep
January 2025
Institut für Chemie, Universität Potsdam, 14476, Potsdam, Germany.
The quantification of different structures, isoforms and types of damage in plasmid DNA is of importance for applications in radiation research, DNA based bio-dosimetry, and pharmaceutical applications such as vaccine development. The standard method for quantitative analysis of plasmid DNA damage such as single-strand breaks (SSB), double-strand breaks (DSB) or various types of base-damage is Agarose gel electrophoresis (AGE). Despite being well established, AGE has various drawbacks in terms of time consuming handling and analysis procedures.
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!