CRISPR in butterflies: An undergraduate lab experience to inactivate wing patterning genes during development

CRISPR is a technique increasingly used in the laboratory for both fundamental and applied research. We designed and implemented a lab experience for undergraduates to carry out CRISPR technology in the lab, and knockout the wing patterning genes optix and WntA in Vanessa cardui butterflies. Students obtained spectacular phenotypic mutants of butterfly wings color and patterns, awakening curiosity about how genomes encode morphology. In addition, students successfully used molecular techniques to genotype and screen wild-type caterpillar larvae and butterflies for CRISPR edits in genes. Student feedback suggests that they experienced a meaningful process of scientific inquiry by carrying out the whole CRISPR workflow process, from the design and delivery of CRISPR components through microinjection of butterfly eggs, the rearing of live animals through their complete life cycle, and molecular and phenotypic analyses of the resulting mutants. We discuss our experience using CRISP genome editing experiments in butterflies to expose students to hands-on research experiences probing gene-to-phenotype relationships in a charismatic and live organism.