Abstract
Phenotype-driven mutagenesis experiments are a powerful approach to identifying novel alleles in a variety of contexts. The traditional disadvantage of this approach has been the subsequent task of identifying the affected locus in the mutants of interest. Recent advances in bioinformatics and sequencing have reduced the burden of cloning these ENU mutants. Here we report our experience with an ENU mutagenesis experiment and the rapid identification of a mutation in a previously known gene. A combination of mapping the mutation with a high-density SNP panel and a candidate gene approach has identified a mutation in collagen type II, alpha I (Col2a1). Col2a1 has previously been studied in the mouse and our mutant phenotype closely resembles mutations made in the Col2a1 locus.
Highlights
Forward genetics in the mouse has been demonstrated to be an efficient tool for taking an unbiased approach to generate novel alleles in genes important for a wide range of functions, including organogenesis
We recently performed an ENU mutagenesis experiment in the mouse as part of our continuing efforts to understand the genetic basis of congenital malformations affecting the forebrain and craniofacial tissues
We began with mutagenized C57BL/6J (B6) G1 males donated from an ENU mutagenesis project at the Jackson Laboratories
Summary
Forward genetics in the mouse has been demonstrated to be an efficient tool for taking an unbiased approach to generate novel alleles in genes important for a wide range of functions, including organogenesis. We harvested the G3 embryos at embryonic day (E) 18.5 with the goal of recovering mutations that might result in severe organogenesis phenotypes which would otherwise be lethal after birth. These skeletal preparations revealed dramatic phenotypes in multiple skeletal elements of mutant embryos as compared to wildtype littermates (Fig. 2C–F).
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