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Updated: May 1, 2026

Mouse Genome Engineering Using Designer Nucleases

Mouse Genome Engineering Using Designer Nucleases

Published on: April 2, 2014

Mouse genome engineering using designer nucleases.

Mario Hermann¹, Tomas Cermak², Daniel F Voytas²

¹Institute of Laboratory Animal Science, University of Zurich.

Journal of Visualized Experiments : Jove

|April 22, 2014

Summary

This summary is machine-generated.

This study presents a protocol for targeted genome modification in mice using transcription activator-like effector nucleases (TALENs) mRNA. This method enables precise genetic alterations without relying on embryonic stem cell technology, advancing disease modeling and therapeutic research.

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Genome Editing in Mammalian Cell Lines using CRISPR-Cas

Genome Editing in Mammalian Cell Lines using CRISPR-Cas

Published on: April 11, 2019

Area of Science:

* Molecular Biology
* Genetics
* Biotechnology

Background:

* Site-specific genome modifications in transgenic mice are crucial for biological research and disease modeling.
* Designer nucleases like ZFNs, TALENs, and CRISPR/Cas9 facilitate targeted genome engineering.
* Current methods often require embryonic stem cell technology, limiting broader application.

Purpose of the Study:

* To provide a protocol for targeted genome modification in mice.
* To demonstrate the efficacy of using transcription activator-like effector nucleases (TALENs) mRNA for genome editing.
* To offer an alternative to embryonic stem cell-based methods for creating genetically modified mice.

Main Methods:

* Identification of designer nuclease target sites within a gene of interest.
* Construction of custom DNA-binding domains for TALENs.
* In vitro transcription of TALEN mRNA and microinjection into fertilized mouse oocytes.

Main Results:

* Successful targeted genome modification achieved through direct injection of TALEN mRNA.
* Demonstrated a viable method for generating genetically modified mice without ES cells.
* Protocol enables rapid and precise genetic alterations in mice.

Conclusions:

* Direct injection of TALEN mRNA is an effective method for targeted genome modification in mice.
* This approach bypasses the need for embryonic stem cell technology, simplifying genome editing.
* The protocol facilitates the generation of transgenic mice for biological studies and therapeutic development.