Date of Award

Spring 5-27-2025

Degree Type

Honors Thesis

Major

Biology

Degree Name

Bachelor of Science

Department

Biology

First Advisor

Dr. Brian Thornton

Second Advisor

Dr. John Gordan

Third Advisor

Dr. Christina Tzagarakis-Foster

Abstract

Fibrolamellar Carcinoma (FLC) is a rare liver cancer, predominantly affecting younger individuals with no history of primary liver disease. As FLC comprises only < 1% of all liver tumors, our understanding of its development and treatment are extremely limited. All clinical cases of FLC contain a specific DNAJB1-PRKACA fusion. The mutation produces an oncogenic form of Protein Kinase A (PKA), known as DNAJ-PKAc, with enhanced binding activity compared to wild-type PKA. Specifically, DNAJ-PKAc interacts with different substrates than wild type PKA, affecting downstream signaling pathways to promote cancer development. However, the genetic dependencies that result from oncogenic DNAJ-PKAc signaling are not known. In this paper, I will show the process of performing a genome-wide CRISPRi screen in AML12DNAJ-PKAc cells to identify genes associated with oncogenic DNAJ-PKAc signaling. I will detail the preliminary benchmarking experiments, the screening process, the preparation of genomic DNA for sequencing, and the ongoing validation of screen results. I anticipate that this project will discover genes whose knockdown inhibits oncogenic DNAJ-PKAc signaling and reduces cell growth exclusively in the presence of PKA. The identified genes could represent potential therapeutic targets for cancer drugs that inhibit FLC progression. However, additional research is necessary to characterize their interactions with DNAJ-PKAc and their specific role in FLC development.

Download

Share

COinS