Sean Wollard

Date of Award

2021

Document Type

Thesis

Degree Name

Bachelors

Department

Natural Sciences

First Advisor

Walstrom, Katherine

Area of Concentration

Chemistry

Abstract

The insulin/insulin-like growth factor (IIS) pathway and adenosine deaminases acting on RNA (ADARs) have both been found to be highly conserved in eukaryotic species and in their importance to aging and lifespan. The proposed research suggests that an up stream regulator of the IIS pathway, ASNA-1 ATPase, which is also highly conserved, may be regulated by the ADAR, ADR-2, in Caenorhabditis elegans (C. elegans). This would provide a mechanism in which ADARs act as regulators of the IIS pathway, which has been hypothesized by previous research. This research utilizes C. elegans adr-2/null mutants and RNAi suppression of the gene coding for ASNA-1 ATPase, asna-1, along with qPCR analysis of the down stream targets of ASNA-1 ATPase in the IIS pathway, daf-28 and daf-16, in order to determine if the presence of ADR-2 has an effect on the function of ASNA-1 ATPase on the IIS pathway. RNAi suppression of asna-1 was performed by feeding the worms E. coli with an asna-1/RNAi plasmid vector on NGM plates utilizing a “half- and-half”, half wild-type E. coli and half asna-1/RNAi, lawn on the plates ensured that the adr-2/null;asna-1/RNAi worms were able to survive long enough for qPCR harvesting. Though the experiment was cut-short due to the COVID-19 outbreak, the research found that adr-2 and asna-1 could likely be absent at the same time in L1 phase C. elegans. The possible results of the uncompleted experiments were also proposed.

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