Graduation Year

2026

Document Type

Campus Only Senior Thesis

Department

Biochemistry

Terms of Use & License Information

Terms of Use for work posted in Scholarship@Claremont.

Rights Information

© 2025 Simran K Sethi

Abstract

POU2AF2, a transcriptional co-factor essential for tuft cell differentiation and epithelial immune signaling, has recently emerged as a potential regulator of colorectal cancer (CRC) biology. Tuft-cell–like transcriptional programs, characterized by POU2F3 and POU2AF2, define a distinct subset of CRCs with unique therapeutic vulnerabilities. However, the functional contribution of POU2AF2 to tumor behavior remains poorly understood. This study aims to characterize early phenotypic and molecular consequences of CRISPR-mediated POU2AF2 knockout in human CRC cell lines. Preliminary growth analyses were performed to establish baseline proliferation rates, followed by gPCR amplification and sequencing to confirm the presence of POU2AF2 target sequences across the CRC cell lines. Once confirmed, PX459-based CRISPR plasmids were transfected into the cell lines. Initial gPCR screens confirmed successful genomic DNA isolation and amplification. Although pilot transfection efficiency was low in HCT-116 and HT-29 cells, the puromycin selection produced viable populations for downstream analysis and RNA sequencing results revealed early changes in differential gene expression. These preliminary findings establish the feasibility of POU2AF2 gene editing in CRC cells and lay foundational work for evaluating how POU2AF2 influences CRC growth and global gene expression. Understanding POU2AF2-dependent pathways may reveal new biomarkers of tumor subtype and uncover transcriptional dependencies that could be leveraged for future targeted therapies in colorectal cancer.

Download

This thesis is restricted to the Claremont Colleges current faculty, students, and staff.

Share

COinS