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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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Updated: Apr 20, 2026

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

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

Published on: August 28, 2018

Using engineered endonucleases to create knockout and knockin zebrafish models.

Victoria M Bedell¹, Stephen C Ekker

¹Department of Biochemistry and Molecular Biology, Mayo Clinic, Guggenheim 1321A, 200 1st Street South, Rochester, MN, 55905, USA.

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

|November 20, 2014

Summary

This summary is machine-generated.

Recent advances in gene-editing technologies like CRISPR/Cas, ZFNs, and TALENs enable targeted knockout and knockin zebrafish. This makes zebrafish a crucial vertebrate model for studying human diseases.

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Last Updated: Apr 20, 2026

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Generation of Maternal Mutants Using zpc:cas9 Knock-in Zebrafish

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Published on: July 22, 2025

Area of Science:

Genetics and Genomics
Developmental Biology
Disease Modeling

Background:

Zebrafish are increasingly utilized as a vertebrate model organism.
Gene-editing technologies have rapidly advanced, offering precise genetic modification capabilities.

Purpose of the Study:

To highlight the advancements in generating targeted knockout and knockin zebrafish.
To present a single-stranded DNA (ssDNA) protocol for creating knockin zebrafish using the TALEN system.

Main Methods:

Utilized zinc finger nucleases (ZFNs), transcription activator-like effector nucleases (TALENs), and CRISPR/Cas systems for targeted gene modification.
Employed a high-efficiency TALEN system and a single-stranded DNA (ssDNA) protocol to generate knockin zebrafish.

Main Results:

Demonstrated the successful creation of targeted knockout zebrafish using ZFNs, TALENs, and CRISPR/Cas.
Successfully generated knockin zebrafish utilizing the TALEN system and the described ssDNA protocol.

Conclusions:

The development of advanced gene-editing tools has significantly enhanced the utility of zebrafish.
Zebrafish serve as a valuable vertebrate model, bridging invertebrate and mammalian systems for human disease research.