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Using ENU mutagenesis for phenotype-driven analysis of the mouse.
Rolf W Stottmann1, David R Beier
1Division of Genetics, Department of Medicine, Brigham and Women's Hospital, Harvard Medical School, Boston, Massachusetts, USA.
Methods in Enzymology
|August 12, 2010
Summary
Phenotypic-driven mutagenesis using N-ethyl-N-nitrosourea (ENU) in mice is a powerful tool for discovering gene functions. This method, though labor-intensive, provides unbiased insights into mammalian biology.
Area of Science:
- Genetics
- Developmental Biology
- Mammalian Genetics
Background:
- Phenotypic-driven mutagenesis has a long history in invertebrates.
- N-ethyl-N-nitrosourea (ENU) was identified as a potent mutagen in mice in the 1970s.
- The application of ENU mutagenesis for studying mammalian biology has accelerated with advances in genomic analysis.
Purpose of the Study:
- To provide insight into the biological roles of genes.
- To offer an unbiased approach to studying gene function.
- To discuss practical aspects of implementing phenotype-driven ENU mutagenesis in mice.
Main Methods:
- ENU treatment of mice.
- Phenotypic screening of mutagenized populations.
- Genetic mapping and positional cloning of induced mutations.
- Mutation validation.
Main Results:
- ENU mutagenesis is a highly productive method for generating phenotypic variants in mice.
- Genomic analysis tools facilitate the positional cloning of ENU-induced mutations.
- This approach provides unbiased insights into gene function.
Conclusions:
- Phenotype-driven ENU mutagenesis is a powerful strategy for mammalian genetics research.
- Successful implementation requires careful attention to experimental design and execution.
- This method is essential for understanding gene function without prior assumptions.
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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.
01:16In 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.