Abstract
Neuropeptides and their receptors have diverse and important roles in regulating physiology, including appetite, stress, inflammation and pain, and they function in Drosophila as well as mammals. My longer-term goal is to create a library of CRISPR-mediated knock-outs of all neuropeptide receptor genes in Drosophila, and then develop/use novel methods to create and assay adult cell-specific knock-outs of these genes. I have begun with the adult brain circadian clock network because it is known to be highly regulated by neuropeptides, in part to anticipate environmental changes during the 24 hour cycle. Knocking-out neuropeptide receptors in a neuron-specific manner may therefore provide important insights into communication within the circadian network. To assess our methods and strategies, I began by generating cell-specific knockouts of the core clock proteins Period (PER) and Timeless (TIM) to show that the tRNA-sgRNA CRISPR-Cas9 system works efficiently to disrupt these genes within the clock neural network. The results also have some surprising features. I have just begun to assay the application of this technique to neuropeptide receptors but have already confirmed that the short neuropeptide F receptor (sNPFR) positively and strongly regulates sleep. Further investigation promises to provide a deeper understanding of how neuropeptides function in neuronal communication and how they influence health and disease.