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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: May 2, 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 knockout mice using engineered nucleases.

Young Hoon Sung¹, Young Jin¹, Seokjoong Kim²

¹Department of Biochemistry, College of Life Science and Biotechnology, Laboratory Animal Research Center, Yonsei University, Seoul 120-749, Republic of Korea.

Methods (San Diego, Calif.)

|February 25, 2014

Summary

This summary is machine-generated.

Engineered nucleases like TALENs and RNA-guided endonucleases (RGENs) offer an efficient method for creating gene knockout and knockin mice. These tools induce targeted DNA breaks in early embryos, facilitating precise genetic modifications.

Keywords:

Engineered nuclease Knockout mouse Microinjection NHEJ RGEN TALEN ZFN

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Last Updated: May 2, 2026

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Mouse Genome Engineering Using Designer Nucleases

Mouse Genome Engineering Using Designer Nucleases

Published on: April 2, 2014

Area of Science:

Molecular Biology
Genetics
Developmental Biology

Background:

Conventional gene targeting in mouse embryonic stem (ES) cells is being supplanted by engineered nucleases.
Engineered nucleases induce targeted double-strand breaks (DSBs) at specific genomic loci.
Mutations are introduced via error-prone or error-free DNA repair, leading to gene function alteration.

Purpose of the Study:

To provide experimentally validated guidelines for generating mutant mice using engineered nucleases.
To detail methods for non-homologous end-joining (NHEJ)-mediated mutations.
To focus on transcription activator-like effector nucleases (TALENs) and RNA-guided endonucleases (RGENs).

Main Methods:

Microinjection of TALENs and RGENs into one-cell stage mouse embryos (cytoplasm or pronucleus).
Utilizing non-homologous end-joining (NHEJ) repair pathway for mutation induction.
Application of engineered nucleases for generating gene knockout and knockin mice.

Main Results:

Successful generation of gene knockout and knockin mice using TALENs and RGENs.
Efficient introduction of mutations through targeted DSBs and subsequent DNA repair.
Validated guidelines for practical application in mouse mutagenesis.

Conclusions:

Engineered nucleases provide an efficient alternative to traditional gene targeting methods.
TALENs and RGENs are effective tools for rapid generation of genetically modified mice.
The provided guidelines facilitate the creation of mutant mice for research purposes.