Investigating the Molecular Mechanism of GLUT4 Trafficking

Undergraduate Honors Thesis

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Citeable URL: https://scholar.colorado.edu/concern/undergraduate_honors_theses/8336h241v

Abstract

The insulin-stimulated glucose transporter GLUT4 is essential to maintaining metabolic homeostasis in mammalian cells. Regulated trafficking of GLUT4 to and from the plasma membrane relies on a complex molecular mechanism integrating insulin signaling and vesicle transport. Understanding this pathway becomes increasingly crucial as Type II Diabetes, a disease characterized by insulin resistance, becomes more prevalent in the population. Despite decades of research, the molecular mechanism of GLUT4 trafficking is not yet fully understood. CRISPR-Cas9 allows for the ablation of individual genes so their role in GLUT4 trafficking can be investigated. A flow cytometry-based assay is used in this study to assess the effects of gene knockout. This project describes the creation of six knockout cell lines to study the role of EXOC3, EXOC7, OSBPL8, OSBPL10, tomosyn-1, and tomosyn-2 in GLUT4 trafficking. Based on the results of these experiments, it appears EXOC7 acts as a positive regulator of GLUT4 translocation while OSBPL8, OSBPL10, tomosyn-1, and tomosyn-2 act as negative regulators.

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