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In-vitro Mutagenesis

In-vitro Mutagenesis

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.

In vitro Mutagenesis

In vitro Mutagenesis

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.

Conservative Site-specific Recombination and Phase Variation

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.
The recognition sites for Cre recombinase called LoxP...

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Related Experiment Video

Updated: Jun 11, 2026

Site-Directed Mutagenesis for In Vitro and In Vivo Experiments Exemplified with RNA Interactions in Escherichia Coli

Site-Directed Mutagenesis for In Vitro and In Vivo Experiments Exemplified with RNA Interactions in Escherichia Coli

Published on: February 5, 2019

Methods for gene cloning and targeted mutagenesis.

Glen P Carter¹, Dena Lyras, Rachael Poon

¹Department of Microbiology, Monash University, Clayton, Australia.

Methods in Molecular Biology (Clifton, N.J.)

|July 3, 2010

Summary

Clostridium difficile causes intestinal diseases, with hypervirulent strains increasing mortality. New genetic tools now allow molecular study of this pathogen, aiding therapeutic development.

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Last Updated: Jun 11, 2026

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Area of Science:

Microbiology
Infectious Diseases
Molecular Biology

Background:

Clostridium difficile is a bacterium responsible for intestinal diseases, known as Clostridium difficile-associated disease (CDAD).
Emerging hypervirulent strains of C. difficile are linked to higher mortality and disease severity in humans.
Understanding C. difficile at a molecular level is crucial for developing effective treatments.

Purpose of the Study:

To investigate the molecular mechanisms of C. difficile pathogenesis.
To facilitate the rational design of novel therapeutics against C. difficile infections.
To overcome historical challenges in the genetic manipulation of C. difficile.

Main Methods:

Development of methods for introducing plasmid DNA into C. difficile.
Establishment of techniques for generating isogenic mutants.
Application of genetic manipulation tools for studying gene function.

Main Results:

Recent advancements have enabled plasmid DNA introduction into C. difficile.
Effective generation of isogenic mutants is now possible.
These genetic tools are vital for studying gene function in C. difficile.

Conclusions:

The development of genetic manipulation methods has significantly advanced C. difficile research.
These tools are essential for understanding C. difficile virulence and for designing new therapies.
Further molecular studies are critical for combating C. difficile-associated disease.