Researcher(s)
- Jane Oommen, Biological Sciences, University of Delaware
Faculty Mentor(s)
- Aimee Jaramillo-Lambert, Biology, University of Delaware
Abstract
Generating tools to map the localization of EGG–3 and PERM-4 in the oocyte membrane in C. elegans.
Jane Oommen, Aimee Jaramillo-Lambert
EGG-1 and EGG-2, oocyte surface proteins, are key components of egg activation and eggshell structure. Defects in EGG-1 and EGG-2 lead to polyspermy and sterility. Previous research from our lab has shown that these proteins are also required for the proper localization of eggshell and oocyte membrane proteins such as CHS-1, CBD-1, and MBK-2. However, EGG-3 (part of EGG complex) and PERM-4 (part of the PERM complex) are additional proteins involved in egg activation and vitelline layer construction of the C. elegans egg, respectively. The localization of these proteins has not yet been explored in the context of an EGG-1/EGG-2 deletion. To visualize the localization of EGG-3, I aim to generate a strain of C. elegans expressing EGG-3 with a green fluorescent protein (GFP) fused to its N-terminus. Proteins tagged with GFP can fluoresce green and can be visualized with a fluorescence microscope. To create this strain, I injected a CRISPR-Cas9 complex into the parent worm and tested its progeny for the desired mutation. To visualize PERM-4 localization, I performed a genetic cross and phenotyped and genotyped the F2 progeny to generate a strain with both perm-4::mScarlet and egg-1/2Δ/qC1. mScarlet tags PERM-4 with a red fluorescent protein, and qC1 is used as a balancer to prevent loss of the egg-1/2 deletion. With these new strains, I plan to use a combination of microscopy and live imaging to track the interactions between these and neighboring proteins. In addition to the original goal, I also used AlphaFold3 to model protein interactions and label various domains in these complexes. Through these efforts, we can further understand the roles of EGG-1 and EGG-2, specifically exploring their effects on EGG-3 and PERM-4 localization in oocyte membrane formation and egg activation.