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Mouse mutants from chemically mutagenized embryonic stem cells.

R J Munroe¹, R A Bergstrom, Q Y Zheng

¹The Jackson Laboratory, Bar Harbor, Maine, USA.

Nature Genetics

|March 4, 2000

Summary

This summary is machine-generated.

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Researchers developed a new method using ethyl methanesulfonate (EMS) to create mouse mutants from mutagenized embryonic stem (ES) cells. This approach enables efficient generation and screening of mutations for gene function studies.

Area of Science:

Genetics
Developmental Biology
Genomics

Background:

Large-scale generation of mouse mutants is crucial for characterizing human gene functions globally.
Conventional N-ethyl-N-nitrosourea (ENU) mutagenesis faces limitations in monitoring efficiency, strain variation, and requires extensive husbandry.
New methods are needed to overcome these obstacles for efficient mouse mutant generation.

Purpose of the Study:

To develop novel protocols for generating mouse mutants using mutagenized embryonic stem (ES) cells.
To overcome limitations associated with conventional germ-cell mutagenesis.

Main Methods:

Protocols were devised to generate germline chimaeric mice from embryonic stem (ES) cells treated with ethyl methanesulfonate (EMS).
Mutation rates were assessed, with up to 1 in 1,200 mutations observed at the Hprt locus.

Related Experiment Videos

Mutation spectrum analysis was performed for EMS and ICR191.

Main Results:

Germline chimaeras were successfully generated from EMS-mutagenized ES cells.
EMS-induced mutations were transmitted, affecting various processes including limb development, hair growth, hearing, and gametogenesis.
The mutation spectrum induced by EMS was consistent with other mammalian cell studies.

Conclusions:

The developed ES cell-based mutagenesis strategy offers advantages over traditional methods.
This technology allows for shortened breeding schemes and direct screening of mutant phenotypes in ES cells or derivatives.
It provides a powerful tool for large-scale functional genomics and the study of gene function.