Date of Project

5-2-2024

Document Type

Honors Thesis

School Name

College of Arts and Sciences

Department

Biology

Major Advisor

Dr. Mary Kroetz

Second Advisor

Dr. Gail Ramirez-Icaza

Abstract

All living things require some form of genetic regulation so that gene products are correctly produced and maintained. One significant form of post-transcriptional gene expression is RNA silencing, a biological mechanism in which double stranded RNA molecules inhibit gene expression by blocking translation of targeted mRNA molecules. Due to its extensive applications in biotechnology, it’s been adapted as an experimental technique referred to as RNAi. This technique is temporary, transient, and easy to use. A new form of RNAi, referred to as the “Flexon” technique, is a novel approach that provides a method for post-transcriptional gene regulation that allows for tissue specific RNAi in the gonads of nematodes. In C. elegans, the Flexon cassette is inserted into the genome along with a Cre driver, allowing for the study of genes essential for viability but also are suspected to have a significant role in gonadogenesis when Cre-recombinase is expressed. When expressed, excision of the Flexon results, causing the restoration of RNAi in the gonads. The Flexon cassette can also be utilized for tissues beyond the gonad in RNAi. To assess the efficacy of this technique, we generated a new strain of C. elegans that contains the Flexon cassette, a Cre driver, and a him-8 nondisjunction mutation. This results in the generation of male C. elegans containing the Flexon and the gonad specific Cre driver. Using this strain, the efficacy of the Flexon technique will be evaluated through inhibition of a gene known to play a role in the development of the male gonad of C. elegans that is also important for embryonic and larval viability. The gene, known as hnd-1, will be inhibited in male C. elegans, and we will observe for visually apparent deformities in the gonad but in other tissues in the animal. Afterwards, additional genes will be tested.

Download

Included in

Genetics Commons

Share

COinS