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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.

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Tamoxifen feeding method is suitable for efficient conditional knockout.

Kumiko Yoshinobu¹, Masatake Araki¹, Ayaka Morita¹

¹Institute of Resource Development and Analysis, Kumamoto University, 2-2-1 Honjo, Chuo-ku, Kumamoto 860-0811, Japan.

Experimental Animals

|October 15, 2020

Summary

Researchers developed a new method for administering tamoxifen to mice using powdered food. This technique achieves efficient genetic recombination without harming the animals, offering a safer alternative for generating conditional knockout models.

Keywords:

Cre conditional knockout feeding method tamoxifen

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Area of Science:

Genetics
Molecular Biology
Animal Models

Background:

Conditional knockout mice are essential for studying gene function.
Tamoxifen administration is commonly used to induce Cre-mediated recombination in these models.
Current administration methods, like intraperitoneal injection, can be invasive and stressful for mice.

Purpose of the Study:

To develop and evaluate a novel, less invasive method for tamoxifen administration in mice.
To assess the efficiency of tamoxifen-induced Cre recombination using the new method.
To compare the safety and efficacy of the novel method against traditional administration routes.

Main Methods:

A tamoxifen-inducible Cre-driver mouse line was utilized for ubiquitous recombination studies.
Tamoxifen was administered to mice mixed with powdered food.
Cre recombination efficiency was assessed via X-gal staining intensity.
The method's applicability to prenatal and fetal periods was investigated.

Main Results:

The powdered food method successfully induced efficient Cre recombination throughout the whole body.
X-gal staining intensity was comparable to that achieved with intraperitoneal tamoxifen administration.
This feeding method proved to be non-injurious to the mice.
The method demonstrated efficacy for recombination during prenatal and fetal developmental stages.

Conclusions:

Administering tamoxifen via powdered food is an effective and safer alternative to traditional methods.
This approach minimizes animal distress and potential injury.
The feeding method is suitable for inducing recombination across various developmental stages, including prenatal.
Researchers are encouraged to adopt this improved tamoxifen delivery system for generating conditional knockout mice.