Generating a novel fly line to quantify cellular oxidative stress on developing Drosophila melanogaster
Area of Study
Science and Mathematics
The redox condition of the cell, the ratio of reactive oxygen species to reducing agents present, can indicate the likelihood of oncogenesis. There are several genetic pathways present, such as the Nrf2 pathway in humans, which moderate the level of reactive oxygen species (ROS) within the cell. The Nrf2 pathway is an orthologue to the cncC pathway in Drosophila melanogaster, making Drosophila an ideal model organism to study redox pathways in the cell. Previous work has shown that cncC alterations can alter oncogene levels in Drosophila, however it hasn’t been shown that this manipulation truly alters ROS levels within the cell.
To solve this, a fly line was created which could indicate and quantify ROS levels via fluorescence microscopy. This was done through use of the UAS-GAL4 system; a system which allows for the introduction and expression of desired genes within the Drosophila melanogaster genome. For this desired purpose, both the UAS-GAL4 indicator and ROS GFP reporter need to be on the same chromosome, so recombination was necessary for the success of this introduction. Through several rounds of genetic crosses, a fly line was established with a UAS-GAL4 indicator and a ROS GFP reporter on the same chromosome within the selected flies. Recombination occurred in 26% of 250 offspring. This recombination will allow quantification of ROS in developing wings, guts, and testes via GFP fluorescence.
Washburn, Hanna, "Generating a novel fly line to quantify cellular oxidative stress on developing Drosophila melanogaster" (2022). Summer Research. 444.
University of Puget Sound