Date of Project

4-28-2026

Document Type

Honors Thesis

School Name

College of Arts and Sciences

Department

Biology

Major Advisor

Mary B Kroetz

Second Advisor

Josef J Jareczek

Abstract

To elucidate the molecular role of essential genes in the gonadal development of Caenorhabditis elegans (C. elegans) it is necessary to utilize specialized molecular techniques such that the phenotypic effects of gene knockdown can be studied while the viability of the organism is preserved. Shaffer and Greenwald (2022) developed the floxed exon (flexon), a tool that improves upon previous approaches to spatiotemporal control of gene expression. The flexon subunit is made up of an artificial exon with a stop cassette flanked by artificial introns which, when inserted into a gene of interest, prevents the expression of that gene. When Cre recombinase is expressed in the presence of the flexon, the stop cassette is excised, and the gene of interest can be expressed. A tissue specific promoter can be utilized to express Cre recombinase in a tissue specific manner. In this study, a gonad specific promoter was utilized to excise the flexon and allow RNAi to occur in the gonad. This way, the gene of interest is only expressed in the gonad while the gene’s expression remains abrogated in all other tissues. In order to determine the molecular role of essential genes in the development of the gonad in both male and hermaphrodite C. elegans, a novel strain of worms was produced that contains a mutation resulting in a higher incidence of male progeny in addition to the flexon subunit which disrupts rde-1, a subunit of the RISC complex, and gonad-specific Cre recombinase expression.

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