The use of cultured cell lines can sometimes provide a cost-effective alternative to transgenic animal models for studying gene function and signaling pathways.1 However, validation that a given cell line resembles its primary cell type is critical to confidently draw credible conclusions of any given study. The goal of this project is to create a transgenic clonal population of mouse lens epithelial cells lacking a functional Foxe3 gene. CRISPR/Cas9 gene editing technology will be used to induce a frame-shift mutation in the Foxe3 gene in the 21EM15 lens epithelial cell line. RNA sequencing will be used to compare the Foxe3 null cell line to wild-type 21EM15 cells to gain insights into the genes controlled by FOXE3. This RNA-seq data will also be used to assess congruency between Foxe3 null and wild type 21EM15 cells and existing RNA-seq data from wild-type and Foxe3 null mice.

The goal of this project is to create a Foxe3 null lens epithelial cell line and compare the transcriptome of the 21EM15 cells with and without FOXE3.

•FOXE3 is a transcription factor that controls gene expression in the lens.

•Foxe3 is a single exon gene that encodes a protein of 288 amino acids.

•Mutations in human and mouse Foxe3 lead to cataracts and microphthalmia.

•21EM15 is a mouse lens epithelial cell line that expresses FOXE3.

During my time working in the Robinson lab I have gained many skills that will be beneficial for future use. However critical thinking, communication, and self development are among the most important. In my research critical thinking is extremely important when creating constructs, performing assays, and analyzing RNA sequencing. I have demonstrated strong communication skills through working effectively along side both my lab and faculty members to achieve my lab based goals. Additionally, through completing my project funded by the Undergraduate Research Award as well as working in the lab for 2 years, I have gained confidence in my abilities as an undergraduate researcher. Not only have I expanded my skill set, I have a greater appreciation for the art of genetics.