Bee Project **Retired as of May 2025** Some of our collaborators are moving to a new university, and as such this project is being retired during summer of 2025. It was a great ~4 years of beekeeping and engineering! The Bee Project aims to engineer bee gut bacteria in an effort to better understand the bee genome and how it impacts bee behavior and health, using the FUGUES (Functional Genomics Using Engineered Symbionts) methodology pioneered by the Barrick and Moran labs. This methodology involves engineering bee symbionts, such as Snodgrassella, to produce dsRNA complementary to our gene of interest. In the bee’s cells, this dsRNA will activate an RNA interference (RNAi) response, which degrades any mRNA transcripts that match the dsRNA sequence. This silences the expression of our gene of interest, and the subsequent phenotypic changes can be analyzed to determine genomic function. Our active subprojects: Circadian Rhythms. Bees rely heavily on their diurnal foragers to keep the hive safe from threats and provide the entire colony with food. Changing these diurnal foragers’ circadian rhythms produces readily observable physiological and behavioral effects, which is ideal for demonstrating the utility of FUGUES. By silencing circadian rhythm genes, we can better understand how we can study, alter, and shape the sleep-wake cycles of bees. Cloning. In order to silence specific genes in the bee genome, we harness an innate pathway called RNA interference, by which double-stranded RNA is used to identify specific mRNA sequences for degradation. Through our cloning pipeline, we design and produce plasmids capable of producing dsRNA targeted to key bumblebee circadian rhythm genes. We use Golden Gate Assembly to create these knockdown plasmids in vitro. Electroporation. In order to induce RNA interference in bumblebees, we aim to incorporate our plasmids into bumblebee gut bacteria Snodgrassella communis, then colonize the bee gut with the engineered gut bacteria. Electroporation is a method by which bacteria uptake DNA through temporary pores in their membrane that are created by an electrical pulse. It is currently our technique of choice to place our dsRNA-producing plasmids into S. communis.
