Deconstructing Gene Function through ENU Mutagenesis

Abstract A great majority of genes present in the human genome are also present in the mouse, thus making it an attractive mammalian model organism to study gene function and dysfunction. Over the past few decades, the ability to manipulate the mouse genome has been developed in a variety of ways. A complementary methodology to create mutations in the mouse is to use chemical mutagenesis.N‐ethyl‐N‐Nitrosourea (ENU) is the mutagen of choice for creating random point mutations model organisms. Advances in sequencing technologies have resulted in a rapid identification of the causative mutation. ENU mutagenesis is a powerful hypothesis‐generating approach to create new mouse models through both forward and reverse genetics approaches. Furthermore, the addition of challenges can identify mutations affecting specific pathways, and specific mutant lines or strains can be used to identify modifiers. Key Concepts ENU produces mainly point mutations randomly throughout the genome. ENU mutagenesis can be used in both forward and reverse genetics approaches. ENU phenotype‐driven screens do not require previous knowledge of the gene to create new mouse models or reveal gene function. ENU phenotype‐driven screens can assign functions to specific protein domains. ENU phenotype‐driven screens identify new mouse models by means of phenotyping. High‐throughput sequencing technologies mean that the identification of the mutation underlying an observed phenotype is very rapid. Phenotype‐driven screens can be applied along with specific challenges or mutations to reveal mutations affecting specific pathways or which modify existing phenotypes.