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Related Experiment Video

Updated: Jun 25, 2026

Gene-targeted Random Mutagenesis to Select Heterochromatin-destabilizing Proteasome Mutants in Fission Yeast

Gene-targeted Random Mutagenesis to Select Heterochromatin-destabilizing Proteasome Mutants in Fission Yeast

Published on: May 15, 2018

Gene-trap vectors and mutagenesis.

Silke De-Zolt¹, Joachim Altschmied, Patricia Ruiz

¹Department of Molecular Hematology, University of Frankfurt, Frankfurt am Main, Germany.

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

|March 7, 2009

Summary

Gene trapping efficiently creates mutations in mouse embryonic stem cells (ESCs) to study gene function. This high-throughput method inactivates genes and provides molecular tags for identification, aiding in the creation of mutant mouse lines.

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Area of Science:

Genetics
Molecular Biology
Genomics

Background:

Gene trapping is a powerful technique for generating insertional mutations in mouse embryonic stem cells (ESCs).
This method has been adapted for high-throughput screening to inactivate genes across the mouse genome.
Gene-trap vectors simultaneously disrupt gene expression and facilitate molecular tagging for gene identification.

Purpose of the Study:

To describe the fundamental methodology for inducing and characterizing gene-trap mutations in ESCs.
To highlight the utility of gene trapping for creating comprehensive mutant mouse libraries.
To report the current progress in genome-wide gene inactivation using this approach.

Main Methods:

Utilizing specialized vectors for gene trapping in mouse ESCs.
Implementing high-throughput screening for efficient gene inactivation.
Developing methods for molecular characterization of gene-trap insertions.

Main Results:

Successful induction and characterization of gene-trap mutations in ESCs.
Creation of extensive ESC libraries with single-gene disruptions.
Approximately 70% of protein-coding genes in the mouse genome have been disrupted.

Conclusions:

Gene trapping is a viable and effective strategy for large-scale functional genomics in mice.
The described methodology facilitates the generation of valuable resources for studying gene function.
Ongoing efforts aim to achieve genome-wide gene inactivation through continued gene-trap applications.