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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.

Lethal Alleles

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Agouti: A Lethal Allele
Lucien Cuénot discovered lethal alleles in 1905 while studying the inheritance of coat color in mice. The agouti gene is responsible for the color of the coat in mice. This gene codes for an agouti-signaling protein, which is responsible for melanin distribution in mammals. The wild-type allele gives rise to gray-brown coat color in mice, while the mutant allele gives rise to yellow coat color. In addition to coat color, the agouti gene is associated with the yellow...

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Related Experiment Video

Updated: Apr 26, 2026

Improved Genome Editing via Oviductal Nucleic Acids Delivery-based In Vivo Electroporation Technique for Knockout Mice Generation

Improved Genome Editing via Oviductal Nucleic Acids Delivery-based In Vivo Electroporation Technique for Knockout Mice Generation

Published on: August 26, 2025

Generation of conditional knockout mice.

Kazuhito Sakamoto¹, Channabasavaiah B Gurumurthy, Kay-Uwe Wagner

¹Eppley Institute for Research in Cancer and Allied Diseases, University of Nebraska Medical Center, DRCII, Rm. 5031, 985950 Nebraska Medical Center, Omaha, NE, 68198-5950, USA, ksakamoto@unmc.edu.

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

|July 28, 2014

Summary

This summary is machine-generated.

Conditional knockout mice are essential for studying gene function in specific tissues. This chapter details a novel method using lambda Red phage recombination for efficient gene targeting vector construction, simplifying the generation of these valuable research models.

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Published on: July 22, 2025

Area of Science:

Genetics
Molecular Biology
Developmental Biology

Background:

Conditional knockout mouse models enable tissue-specific gene function analysis.
Traditional methods for generating these models are complex and time-consuming.

Purpose of the Study:

To describe an improved method for generating gene-targeting vectors for conditional knockout mice.
To highlight the advantages of lambda Red phage-based homologous recombination.

Main Methods:

Gene-targeting vector construction using lambda Red phage-based homologous recombination in E. coli.
Transfer of the vector into embryonic stem (ES) cells.
Injection of targeted ES cells into mouse blastocysts.

Main Results:

The described method facilitates the capture of genomic DNA and assembly of gene-targeting vectors.
This approach circumvents the need for restriction sites within the target gene.
Successful generation of conditional knockout mice is achieved through ES cell manipulation and blastocyst injection.

Conclusions:

Lambda Red phage-based homologous recombination offers an efficient and versatile alternative for constructing gene-targeting vectors.
This streamlined approach simplifies the generation of conditional knockout mouse models for biological research.