Abstract
AbstractEstablishing genotype–phenotype relationships is critical for understanding the genetic architecture underlying trait evolution. While functional genetic approaches have long been established in a number of classic model systems, genetic perturbations have historically been challenging in organisms that are used in evolutionary studies. The recent establishment of CRISPR-Cas9-mediated gene editing has now made functional genetic studies possible in a wide range of species. Astyanax mexicanus, a species of freshwater fish which has cave and surface populations, is a powerful system for evolutionary genetic studies. Here, we describe a protocol for using CRISPR-Cas9 to study gene function in A. mexicanus. Specifically, we describe methods for rapid screening of gene function using first generation, CRISPR-Cas9-injected animals, as well as methods for screening for germline transmission to enable the establishment of lines of fish with mutations in genes of interest. Our goal is to provide guidelines for performing functional genetic analyses in A. mexicanus using CRISPR-Cas9 and to highlight the advantages of using this approach.Key words Astyanax mexicanus CavefishCRISPR-Cas9Genome editingFunctional assaysFunctional genomics
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