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Related Concept Videos

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 Repair01:38

Nucleotide Excision Repair

DNA Distortion and Damage
Cells are regularly exposed to mutagens—factors in the environment that can damage DNA and generate mutations. UV radiation is one of the most common mutagens and is estimated to introduce a significant number of changes in DNA. These include bends or kinks in the structure, which can block DNA replication or transcription. If these errors are not fixed, the damage can cause mutations, which in turn can result in cancer or disease depending on which sequences are...
Nucleotide Excision Repair01:08

Nucleotide Excision Repair

Overview
Nucleotide Excision Repair01:08

Nucleotide Excision Repair

Overview
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...
CRISPR01:59

CRISPR

Genome editing technologies allow scientists to modify an organism’s DNA via the addition, removal, or rearrangement of genetic material at specific genomic locations. These types of techniques could potentially be used to cure genetic disorders such as hemophilia and sickle cell anemia. One popular and widely used DNA-editing research tool that could lead to safe and effective cures for genetic disorders is the CRISPR-Cas9 system. CRISPR-Cas9 stands for Clustered Regularly Interspaced Short...

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Related Experiment Video

Updated: May 29, 2026

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

DNA nicks promote efficient and safe targeted gene correction.

Luther Davis1, Nancy Maizels

  • 1Department of Immunology, University of Washington School of Medicine, Seattle, Washington, United States of America.

Plos One
|September 14, 2011
PubMed
Summary

Single-strand breaks (nicks) efficiently correct genes with minimal genomic damage, unlike double-strand breaks. This safer approach offers improved therapeutic potential for targeted gene correction.

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Enhanced Genome Editing with Cas9 Ribonucleoprotein in Diverse Cells and Organisms
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Enhanced Genome Editing with Cas9 Ribonucleoprotein in Diverse Cells and Organisms

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Using Next Generation Sequencing to Identify Mutations Associated with Repair of a CAS9-induced Double Strand Break Near the CD4 Promoter
06:59

Using Next Generation Sequencing to Identify Mutations Associated with Repair of a CAS9-induced Double Strand Break Near the CD4 Promoter

Published on: March 31, 2022

Related Experiment Videos

Last Updated: May 29, 2026

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

Enhanced Genome Editing with Cas9 Ribonucleoprotein in Diverse Cells and Organisms
09:51

Enhanced Genome Editing with Cas9 Ribonucleoprotein in Diverse Cells and Organisms

Published on: May 25, 2018

Using Next Generation Sequencing to Identify Mutations Associated with Repair of a CAS9-induced Double Strand Break Near the CD4 Promoter
06:59

Using Next Generation Sequencing to Identify Mutations Associated with Repair of a CAS9-induced Double Strand Break Near the CD4 Promoter

Published on: March 31, 2022

Area of Science:

  • Molecular Biology
  • Genetics
  • Bioengineering

Background:

  • Targeted gene correction aims to fix mutations using DNA lesions to trigger homologous recombination.
  • Current methods often use double-strand breaks (DSBs), which can cause genomic instability and safety concerns.

Purpose of the Study:

  • To compare the efficacy and safety of single-strand breaks (SSBs, or nicks) versus double-strand breaks (DSBs) for targeted gene correction.
  • To evaluate the potential of nicks as a safer alternative for gene correction therapies.

Main Methods:

  • Inducing both nicks and DSBs at a disease gene locus.
  • Measuring gene correction efficiency via homologous recombination.
  • Assessing local and distal genomic alterations (mutagenesis) following nick or DSB induction.

Main Results:

  • Both nicks and DSBs efficiently promote gene correction.
  • DSBs result in significant local and distant genomic alterations.
  • Nicks lead to gene correction with minimal collateral local mutagenesis, demonstrating a 70-fold increase in efficacy (correction to deletion ratio) compared to DSBs.

Conclusions:

  • Single-strand breaks (nicks) are a safer and more efficacious method for targeted gene correction than double-strand breaks.
  • Nicks minimize unintended genomic alterations, holding significant therapeutic implications for gene correction strategies.
  • Future gene editing tool design, particularly meganucleases, should consider utilizing nicks for enhanced safety and precision.