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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.

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...

Gene Therapy

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Gene therapy is a technique where a gene is inserted into a person’s cells to prevent or treat a serious disease. The added gene may be a healthy version of the gene that is mutated in the patient, or it could be a different gene that inactivates or compensates for the patient’s disease-causing gene. For example, in patients with severe combined immunodeficiency (SCID) due to a mutation in the gene for the enzyme adenosine deaminase, a functioning version of the gene can be inserted. The...

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Related Experiment Video

Updated: Jun 22, 2026

Genetic Manipulation in Δku80 Strains for Functional Genomic Analysis of Toxoplasma gondii

Genetic Manipulation in Δku80 Strains for Functional Genomic Analysis of Toxoplasma gondii

Published on: July 12, 2013

Gene-targeting vectors.

J Simon¹, C Arthur, Victoria A McGuire

¹Transgenic Service, College of Life Sciences, University of Dundee, Dundee, UK.

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

|June 9, 2009

Summary

This summary is machine-generated.

Designing effective gene targeting vectors in mice is crucial for understanding gene function. This guide details essential vector components, design strategies, and conditional targeting methods like Cre/LoxP for successful in vivo research.

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Zinc-finger Nuclease Enhanced Gene Targeting in Human Embryonic Stem Cells

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Published on: August 23, 2014

Area of Science:

Molecular Biology
Genetics
Gene Targeting

Background:

Gene targeting in mice is a powerful technique for in vivo gene function elucidation.
Successful gene targeting relies heavily on the precise design of targeting vectors.

Purpose of the Study:

To provide a comprehensive rationale for designing gene targeting vectors.
To detail essential vector components and considerations for various vector types (deletions, point mutations, insertions).
To describe strategies for conditional gene targeting using Cre/LoxP and Flp/frt systems.

Main Methods:

Review of vector design principles for gene deletions, point mutations, and insertions.
Description of cloning strategies and recombinant screening methods.
Explanation of conditional targeting systems (Cre/LoxP, Flp/frt) and generation of conditional mutations.

Main Results:

Detailed examples of vector designs and cloning strategies are provided.
Effective screening approaches for identifying successful recombinants are outlined.
Methods for generating conditional gene deletions and point mutations are presented, including potential drawbacks.

Conclusions:

Careful design of targeting vectors is fundamental for successful gene function studies in mice.
Conditional targeting systems offer advanced control over gene manipulation in vivo.
This chapter serves as a guide for researchers in designing and implementing gene targeting strategies.