Related Experiment Video

Updated: Oct 10, 2025

Characterizing DNA Repair Processes at Transient and Long-lasting Double-strand DNA Breaks by Immunofluorescence Microscopy
08:31

Characterizing DNA Repair Processes at Transient and Long-lasting Double-strand DNA Breaks by Immunofluorescence Microscopy

Published on: June 8, 2018

9.3K

Systematic overview on the most widespread techniques for inducing and visualizing the DNA double-strand breaks

Ivett Berzsenyi1, Vasiliki Pantazi1, Barbara N Borsos1

  • 1Institute of Pathology, Albert Szent-Györgyi Medical School, University of Szeged, 1 Állomás Street H-6725, Szeged, Hungary.

Mutation Research. Reviews in Mutation Research
|December 11, 2021
PubMed

Insights

Cells use DNA repair mechanisms to prevent cancer. This review details methods for inducing and tracking DNA double-strand breaks (DSBs) and their repair in human cells, aiding genome integrity research.

Area of Science:

  • Molecular Biology
  • Genetics
  • Cell Biology

Background:

  • DNA double-strand breaks (DSBs) are a significant cause of cancer initiation.
  • Cellular DNA repair mechanisms are crucial for maintaining genome integrity and function.
  • Dysregulation of DNA repair pathways can lead to genome instability and disease.

Purpose of the Study:

  • To review and summarize methods for studying DNA double-strand break (DSB) repair.
  • To highlight techniques for site-specific DSB induction and repair tracking in human cells.
  • To discuss the advantages and disadvantages of current DSB research methodologies.

Main Methods:

  • Site-specific DSB induction using restriction endonucleases, CRISPR-Cas9, and laser microirradiation.
  • Visualization and kinetic tracking of DSB repair via fluorescence, confocal, and super-resolution microscopy.
  • Employing chromatin immunoprecipitation (ChIP), DSB-labeling, and sequencing techniques for repair analysis.

Main Results:

  • The review provides a comprehensive overview of established methods for DSB research.
  • It compares the strengths and limitations of various DSB induction and repair monitoring techniques.
  • Key approaches for site-specific DSB generation and subsequent repair kinetics are detailed.

Conclusions:

  • Understanding DSB repair mechanisms is vital for combating cancer.
  • The reviewed methods offer powerful tools for advancing research in DNA repair and genome stability.
  • This summary aids researchers in selecting appropriate techniques for their specific DSB studies.
Keywords:
Chromatin immunoprecipitationDNA repairFluorescence microscopyImmunostainingLaser microirradiationSite-specific DSBs

More Related Videos

Visualization of miniSOG Tagged DNA Repair Proteins in Combination with Electron Spectroscopic Imaging ESI13:06

Visualization of miniSOG Tagged DNA Repair Proteins in Combination with Electron Spectroscopic Imaging ESI

Published on: September 24, 2015

10.2K
Visualizing and Quantifying Endonuclease-Based Site-Specific DNA Damage10:59

Visualizing and Quantifying Endonuclease-Based Site-Specific DNA Damage

Published on: August 21, 2021

3.7K

Related Concept Videos

Fixing Double-strand Breaks02:04

Fixing Double-strand Breaks

The double-stranded structure of DNA has two major advantages. First, it serves as a safe repository of genetic information where one strand serves as the back-up in case the other strand is damaged. Second, the double-helical structure can be wrapped around proteins called histones to form nucleosomes, which can then be tightly wound to form chromosomes. This way, DNA chains up to 2 inches long can be contained within microscopic structures in a cell. A double-stranded break not only damages...
13.1K
Homologous Recombination02:31

Homologous Recombination

The basic reaction of homologous recombination (HR) involves two chromatids that contain DNA sequences sharing a significant stretch of identity. One of these sequences uses a strand from another as a template to synthesize DNA in an enzyme-catalyzed reaction. The final product is a novel amalgamation of the two substrates. To ensure an accurate recombination of sequences, HR is restricted to the S and G2 phases of the cell cycle. At these stages, the DNA has been replicated already and the...
55.3K
Overview of DNA Repair02:25

Overview of DNA Repair

In order to be passed through generations, genomic DNA must be undamaged and error-free. However, every day, DNA in a cell undergoes several thousand to a million damaging events by natural causes and external factors. Ionizing radiation such as UV rays, free radicals produced during cellular respiration, and hydrolytic damage from metabolic reactions can alter the structure of DNA. Damages caused include single-base alteration, base dimerization, chain breaks, and cross-linkage.
Chemically...
32.1K
Nucleotide Excision Repair01:08

Nucleotide Excision Repair

Overview
37.9K
The DNA Replication Fork01:02

The DNA Replication Fork

An organism’s genome needs to be duplicated in an efficient and error-free manner for its growth and survival. The replication fork is a Y-shaped active region where two strands of DNA are separated and replicated continuously. The coupling of DNA unzipping and complementary strand synthesis is a characteristic feature of a replication fork.   Organisms with small circular DNA, such as E. coli, often have a single origin of replication; therefore, they have only two replication...
37.6K
Long-patch Base Excision Repair01:02

Long-patch Base Excision Repair

Since the discovery of the two BER pathways, there has been a debate about how a cell chooses one pathway over the other and the factors determining this selection. Numerous in vitro experiments have pointed out multiple determinants for the sub-pathway selection. These are:
7.3K