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Related Concept Videos

Conservative Site-specific Recombination and Phase Variation02:53

Conservative Site-specific Recombination and Phase Variation

Because the DNA segments are cut and reorganized in a direction-specific manner, site-specific recombination has emerged as an efficient genetic engineering technique. Flippase and Cyclization recombinases or Flp and Cre, respectively, are two members of the tyrosine recombinase family derived from bacteriophages, that are used to mediate site-specific DNA insertions, deletions, and targeted expression of proteins in mammalian cell lines.
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In-vitro Mutagenesis01:16

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To learn more about the function of a gene, researchers can observe what happens when the gene is inactivated or “knocked out,” by creating genetically engineered knockout animals. Knockout mice have been particularly useful as models for human diseases such as cancer, Parkinson’s disease, and diabetes.

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Related Experiment Video

Updated: May 14, 2026

In Vitro Directed Evolution of a Restriction Endonuclease with More Stringent Specificity
09:16

In Vitro Directed Evolution of a Restriction Endonuclease with More Stringent Specificity

Published on: March 25, 2020

High frequency targeted mutagenesis using engineered endonucleases and DNA-end processing enzymes.

Fabien Delacôte1, Christophe Perez, Valérie Guyot

  • 1Cellectis SA, Paris, France.

Plos One
|January 30, 2013
PubMed
Summary
This summary is machine-generated.

This study enhances gene inactivation by combining engineered nucleases with DNA-end processing enzymes. This method significantly boosts targeted mutagenesis efficiency up to 30-fold in human cells.

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Last Updated: May 14, 2026

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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

Published on: August 25, 2017

Area of Science:

  • Molecular Biology
  • Gene Editing
  • Genetics

Background:

  • Targeting DNA double-strand breaks is crucial for gene inactivation.
  • Non-Homologous End joining (NHEJ) pathways facilitate targeted mutagenesis without repair plasmids.
  • Engineered nucleases can produce precise DNA re-ligation, limiting mutagenesis efficiency.

Purpose of the Study:

  • To develop a robust and efficient method for increasing targeted mutagenesis frequency.
  • To control the nature of mutagenic events in human primary cells.
  • To overcome limitations of precise DNA re-ligation in gene editing.

Main Methods:

  • Combining engineered endonucleases with DNA-end processing enzymes.
  • Utilizing meganucleases in conjunction with DNA-end processing enzymes.
  • Application in human primary cells for gene inactivation studies.

Main Results:

  • Achieved a significant increase in targeted mutagenesis frequency, up to 30-fold.
  • Demonstrated control over the types of mutagenic events induced.
  • Successfully applied the method in human primary cells.

Conclusions:

  • The combined approach of engineered endonucleases and DNA-end processing enzymes offers a powerful strategy for gene inactivation.
  • This method significantly enhances the efficiency and controllability of targeted mutagenesis.
  • The findings provide a robust tool for genetic research and therapeutic applications in human cells.