1) The big questions Convergent evolution is a natural experiment in repeated evolution of similar phenotypes, offering unique insights into the evolutionary process. When similar patterns evolve in different lineages, to what extent are the same molecular mechanisms deployed? Are the same regulatory changes co-opted into generating convergent phenotypes? How can evolutionary change at a single locus regulate complex patterning changes during rapid evolution? What are the precise genetic changes necessary for the evolution of a new developmental pattern? Heliconius butterflies are an excellent system to address these questions. 2) The background Many tropical butterflies have mimetic wing patterns to warn predators of their toxicity, and these have become an excellent system in which to understand the molecular basis for convergence and diversification, and make the link between natural selection in the wild and evolution in the genome. Here we will study the molecular basis for pattern convergence in tropical Heliconius butterflies. Genetic mapping and gene expression experiments have identified a simple system of three genetic loci that control the complex diversification in wing patterning seen in Heliconius. One of these loci regulates yellow pattern elements and the same genomic locus is also involved in wing patterning of both the peppered moth, Biston betularia, and the butterfly Bicyclus anynana, suggesting an ancient shared patterning system in butterflies and moths. Patterns of expression and genetic data from natural populations, suggest that cortex is the functional gene at this locus, although expression data also point to involvement of other linked genes. We will apply recently developed CRISPR/Cas9 methods for gene knockouts to investigate the molecular basis for pattern convergence between species of Heliconius butterflies. 3) Objectives and expected results We will test the frequency with which similar patterns evolving in mimetic butterflies use the same genes. At a closer resolution, we will also test whether those genes are controlled by the same regulatory switches to turn them on and off, when they control similar patterns. This will test for the repeatability of evolution in different genetic backgrounds. These experiments will involve developing and applying novel gene editing techniques to study patterning in these butterflies, which will set the standard for evolutionary studies in the future. The remarkable patterns of mimicry in these butterflies have long been considered an exemplar of evolution by natural selection, and this project will offer unique new insights into the molecular mechanisms that produce such strikingly similar patterns in so many different species.