Date of Project

12-12-2024

Document Type

Capstone

School Name

College of Arts and Sciences

Department

Biology

Major Advisor

Mary Kroetz

Second Advisor

Josef Jareczek

Abstract

Caenorhabditis elegans are model organisms frequently studied to better understand biological processes. They have short life spans and produce a large number of progeny. Additionally, many of their molecular development processes can be found in human, providing insight into cell death and cell fate. In this project, our research aims to create a triple mutant strain to test the importance of the pig-1 gene in the gonad of C. elegans. This strain carries the following alleles: ckb-3::degron; him-8; gfp::pig-1. The pig-1 gene has been previously studied in neuroblast cells in C. elegans and is required for asymmetrical division, resulting in one neural precursor and one apoptotic cell. The gfp::pig-1 strain was made by a previous undergraduate student Peyton Young. The him-8 gene, when mutant, leads to males making up approximately ~30% of the population. In comparison, males are approximately 0.2%-0.5% of the wild-type population. mRNA transcripts of the pig-1 gene have previously been shown to be enriched in the male gonads. Therefore, the him-8 mutant allele will allow us to phenotypically study the gonads of the male animals. Finally, ckb-3::degron is used to degrade GFP and proteins fused to it. The ckb-3::degron gene allows for the degradation of only gonadal GFP by ubiquitination because the ckb-3 promoter drives the degron to only be expressed in the gonad. The pig-1 gene is essential for the viability of the animal in other tissues and would lead to the animal's death if removed from the whole animal. The triple mutant strain will allow us to observe what will happen to gonad development and function if pig-1 is removed from the gonad in both male and hermaphrodite worms. Therefore, I will analyze the triple mutant worm to see how the gonad is affected by the absence of the PIG-1 protein. This will be done by observing the triple mutant worms and noting any physical or behavioral abnormalities compared to worms containing gfp::pig-1, but not the ckb-3::degron.

Download

Share

COinS