EDGE CT: Precise genome editing in a lepidopteran insect tailored for stable transformation

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One out of ten species known to humans are moths and butterflies of the Lepidoptera order. Although this biodiversity provides new opportunities to understand the genetics of those insects, experimenters have been limited by the lack of a tractable laboratory system for routine genetic analysis. In this Enabling Discovery through Genomic Tools project, the team of investigators develop the Indian-meal moth as a laboratory system for discovery in lepidopteran genomes. This species is highly fertile, hardy, economical to maintain, allowing the development of moth lines embedded with genetic tools. In particular, the investigators optimize cutting-edge genome editing techniques for marking cells and tissues of interest. These efforts will open new avenues of research about the physiology, immunology, reproduction, developmental genetics, and evolution of an insect order of enormous diversity that includes numerous species of economic and ecological importance. To maximize worldwide impact, this project includes the training of young scientists, and the assemblage of a collaborative community of genome editors working with a pest that infests grain storage units and households alike all over the world.To address the lack of model system for routine functional genomics in Lepidoptera, this Enabling Discovery through Genomic Tools project will develop and disseminate tissue-specific assays and genome editing standards fro this lineage using the mealmoth Plodia interpunctella. In addition of being an important pest of stored food across the world, this organism is tailored for genome editing due to a unique suit of reproductive and rearing features (e.g., ease and low-cost of maintenance, injectable eggs, resistance to inbreeding, successful CRISPR targeting), an available genome sequence, and translucent stages and tissues that facilitate the screening of fluorescent markers. In particular, variations of CRISPR Homology-Directed Repair (HDR) is optimized using a dominant rescue assay in which thousands of injected individuals are screened for the targeted repair of a recessive mutation. Optimization assays are designed to yield meaningful comparisons of the in vivo performances of transformation methods for short allele replacements and long DNA knock-ins, and to catalyze further innovation across other insect emerging systems. Beyond their use in fundamental research, the tools developed here are expected to potentiate the integrated management of crop pests and forest defoliators, and to inform the biology of important food chain components and pollinators. Finally, Plodia itself shows a worldwide distribution and presents a threat to global food security. The low-cost of maintenance and EDGE-derived resources will thus stimulate work in this system at an international scale. As part of this project, the team of investigators organizes symposia at scientific meetings and trains a community of scientists interested in working with Plodia in their laboratories. This project is funded by the Behavioral Systems Cluster of Integrative Organismal Systems in the Directorate for Biological Sciences.This award reflects NSF's statutory mission and has been deemed worthy of support through evaluation using the Foundation's intellectual merit and broader impacts review criteria.