AbstractThe insulin/insulin growth factor signaling (IIS) cascade is one of the most important signaling pathways ininsects. IIS in insects regulate everything from development and growth during the immature stages toreproduction and longevity in adult insects to metabolism and innate immunity during all stages. While insectstypically encode multiple insulin-like peptides (ILPs) these hormones appear to largely activate the IIScascade via a single insulin receptor. This raises a fundamental question as to why multiple ILPs arenecessary. They may be redundant to ensure that mutations in this critical signaling pathway are compensatedfor. They may act in concert to fine tune the physiologies regulated by this cascade. Or they mayindependently control distinct physiological processes. To explore these possibilities we will knock-outindividual ILPs in the mosquito Aedes aegypti using the CRISPR/Cas9 system. Ae. aegypti encodes eightdifferent ILPs and we will knock-out at least three of the most intriguing ones; 1) AaegILP2 an ortholog ofDrosophila DILP2 which has been implicated in regulating lifespan reproduction and development 2)AaegILP6 the only putative insulin growth factor identified in mosquitoes to date and 3) AaegILP3 which hasbeen shown through peptide injections to control metabolism and reproduction. For these three AaegILPknock-outs lines and any others that time and resources allow for we will conduct expression assays invarious tissues various developmental periods and following various physiological events. We will also assessthe impact of the individual AaegILP knock-outs on the following physiologies: growth and developmentlifespan metabolism lifetime fecundity and innate immunity. By the end of this project we will have elucidatedthe basic biological roles of three or more AaegILPs broadening our understanding of key factors regulatingvectorial capacity in this important arboviral vector.