The use of mutagenesis in invertebrates to generate phenotypic variants has a long and productive history. Despite the conclusive demonstration by Russell and colleagues in the 1970s that the chemical N-ethyl-N-nitrosourea (ENU) is a highly effective mutagen in mice, the application of phenotypic-driven mutagenesis as a method to study mammalian biology proceeded slowly. With the development of tools for genomic analysis, the task of positional cloning ENU-induced mutations has become quite feasible, and this approach has recently been widely applied and highly productive. It has specifically lived up to its theoretical utility as means to provide insight into the biological roles of genes that is not biased by presumptions of their function. While the power of this approach is indisputable, the effort necessary for its success remains substantial, requiring careful attention to aspects including ENU treatment, mouse husbandry, screen assay design, genetic mapping, positional cloning, and mutation validation. In this chapter we discuss practical aspects of implementing a phenotype-driven analysis of an ENU-mutagenized mouse population.
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