Date of Award

6-15-2017

Document Type

Thesis - SCU Access Only

Publisher

Santa Clara : Santa Clara University, 2017.

Degree Name

Master of Science (MS)

Department

Bioengineering

First Advisor

Bill Lu

Second Advisor

Zhiwen Zhang

Abstract

Cell derived vesicles known as exosomes are becoming the focus of many in the field of bioengineering. These nano scale vesicles are produced and secreted by cells and allow for the carrying and delivery of bioactive cargo within their lipid bilayer membrane. This study aims to engineer novel exosome transmembrane proteins to display dual purpose tags, enabling exosome visualization and isolation in mammalian cell lines. Three different constructs were created using exosome transmembrane proteins VSVG and CD63 in combination with a fluorescent protein and a 6xHis tag. Each construct was transfected into a variety of mammalian cell lines to show that the fluorescent protein tag enables exosome visualization and tracking. Co-localization experiments were performed to ensure that the modifications did not affect the specificity of the transmembrane proteins or have a larger effect on the exosomes themselves. Furthermore, SDSPAGE and Western Blot analysis were conducted to characterize the protein size and test the viability of the 6xHis tag. By engineering an exosome transmembrane protein with dual tag reporters, this study emphasizes the potential impact of exosomes in drug delivery, imaging, and therapy.

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