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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...
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...
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...
Translesion DNA Polymerases02:10

Translesion DNA Polymerases

Translesion (TLS) polymerases rescue stalled DNA polymerases at sites of damaged bases by replacing the replicative polymerase and installing a nucleotide across the damaged site. Doing so, TLS allows additional time for the cell to repair the damage before resuming regular DNA replication.
TLS polymerases are found in all three domains of life - archaea, bacteria, and eukaryotes. Of the different classes of TLS polymerases, members of the Y family are fitted with specialized structures that...
Base Excision Repair01:54

Base Excision Repair

One of the common DNA damages is the chemical alteration of single bases by alkylation, oxidation, or deamination. The altered bases cause mispairing and strand breakage during replication. This type of damage causes minimal change to the DNA double helix structure and can be repaired by the base excision repair (BER) pathways. BER corrects damaged DNA sequences by removing the damaged base and restoring the original base sequence using the complementary strand as a template.
The first step of...
Nucleotide Excision Repair01:08

Nucleotide Excision Repair

Overview

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Related Experiment Video

Updated: Jun 10, 2026

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

Targeting DNA double-strand breaks with TAL effector nucleases.

Michelle Christian1, Tomas Cermak, Erin L Doyle

  • 1Department of Plant Pathology, Iowa State University, Ames, Iowa 50011, USA.

Genetics
|July 28, 2010
PubMed
Summary

Researchers developed novel engineered nucleases by combining transcription activator-like effectors (TALEs) with FokI endonuclease. These TALE-nuclease fusions precisely target and cleave specific DNA sequences in vivo for mutagenesis.

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Last Updated: Jun 10, 2026

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

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Published on: June 8, 2018

Visualizing and Quantifying Endonuclease-Based Site-Specific DNA Damage
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A Standard Methodology to Examine On-site Mutagenicity As a Function of Point Mutation Repair Catalyzed by CRISPR/Cas9 and SsODN in Human Cells
10:07

A Standard Methodology to Examine On-site Mutagenicity As a Function of Point Mutation Repair Catalyzed by CRISPR/Cas9 and SsODN in Human Cells

Published on: August 25, 2017

Area of Science:

  • Molecular Biology
  • Genetic Engineering

Background:

  • Engineered nucleases are crucial for targeted DNA modification.
  • Existing methods for site-specific DNA cleavage have limitations.

Purpose of the Study:

  • To create a new class of sequence-specific nucleases.
  • To demonstrate targeted DNA double-strand breaks using novel fusion proteins.

Main Methods:

  • Fusing transcription activator-like effectors (TALEs) to the FokI endonuclease catalytic domain.
  • Designing and testing both native and custom TALE-nuclease constructs.

Main Results:

  • Successfully generated a new class of TALE-nuclease fusions.
  • Demonstrated precise targeting of DNA double-strand breaks at specific genomic loci.
  • Validated the efficacy of both native and custom TALE-nuclease designs.

Conclusions:

  • TALE-nuclease fusions represent a powerful tool for targeted mutagenesis.
  • This technology enables precise genome editing via site-specific DNA cleavage.