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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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Related Experiment Video

Updated: Jul 17, 2025

Gene Trapping Using Gal4 in Zebrafish

Gene Trapping Using Gal4 in Zebrafish

Published on: September 29, 2013

Generation of Conditional Knockout Zebrafish Using an Invertible Gene-Trap Cassette.

Masahito Ogawa¹, Kazu Kikuchi²

¹Department of Cardiac Regeneration Biology , National Cerebral and Cardiovascular Center Research Institute, Osaka, Japan.

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

|September 5, 2023

Summary

Conditional knockout (cKO) technology allows gene inactivation in specific tissues. We present a new protocol for generating cKO zebrafish, enabling precise study of gene function in organ development and regeneration.

Keywords:

Cardiomyocytes Conditional knockout Gene trap Heart regeneration Zebrafish

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Last Updated: Jul 17, 2025

Gene Trapping Using Gal4 in Zebrafish

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Efficient Production and Identification of CRISPR/Cas9-generated Gene Knockouts in the Model System Danio rerio

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Zygote Microinjection for Creating Gene Cassette Knock-in and Flox Alleles in Mice

Zygote Microinjection for Creating Gene Cassette Knock-in and Flox Alleles in Mice

Published on: June 24, 2022

Area of Science:

Genetics
Developmental Biology
Zebrafish Models

Background:

Conditional knockout (cKO) is crucial for studying gene function by enabling targeted gene inactivation.
Previous limitations in zebrafish hindered the development of cKO models, impacting research on organ development and regeneration.
Advances in genome editing have overcome these limitations, allowing for cKO model generation in various organisms.

Purpose of the Study:

To describe a detailed protocol for generating conditional knockout (cKO) zebrafish.
To enable precise temporal and spatial control of gene expression in zebrafish.
To facilitate the study of gene function in zebrafish organ development and regeneration.

Main Methods:

Utilizing Cre-dependent genetic switch systems.
Employing advanced genome editing technologies.
Developing a detailed protocol for cKO zebrafish generation.

Main Results:

Successful generation of conditional knockout (cKO) zebrafish models.
Demonstration of a Cre-dependent genetic switch for temporal gene inactivation.
Establishment of a platform for precise genetic analysis in zebrafish.

Conclusions:

The developed protocol enables the generation of cKO zebrafish, overcoming previous technical barriers.
This advancement provides a powerful tool for investigating gene function in zebrafish development and regeneration.
Conditional knockout zebrafish models are now accessible for a wide range of genetic studies.