Lemmon, Skyler; Chaston, John (Brigham Young University)
Faculty Advisor: Chaston, John (Life Sciences, Plant and Wildlife Sciences)
Over the course of the last year, I have dedicated most of my time in the lab to learning about CRISPR/Cas9 and practicing the laboratory techniques that are necessary to make genetic changes in Drosophila melanogaster. Here I aim to expand on that expertise by applying CRISPR to study a genetic question: how the microbial composition of the D. melanogaster microbiome is affected by the modification of 4 specifically selected genes in flies from Florida and Maine. For each of the selected genes, the Florida fly allele will be put into the Maine fly genome and the Maine fly allele will be put into the Florida fly genome. The microbiome composition of these two new flies will be compared against the original lines in a factorial design. Embryos will be injected with the necessary plasmids for a double-stranded cut to take place. After injection, homology dependent repair that will incorporate the new allele. Sanger sequencing will be used to screen for successful knock-in of the allele. Finally, the concentrations of each type of bacteria found in the microbiota of the flies will be measured and compared against the flies from which the allele came from.