Date of Award
8-16-2024
Document Type
Open Access Thesis
Department
Chemistry and Biochemistry
First Advisor
Nicholas Truex
Abstract
Cytokines are small immune signaling proteins responsible for cellular communication and their levels are tightly regulated throughout immune recognition and defense. Interferon gamma is a key cytokine responsible for upregulation of antigen presentation and supports the differentiation of pro-inflammatory effector cells. Due to its role in regulation, excess interferon gamma is cleared from the body limiting the success of interferon gamma immunotherapies. This study reports the design and validation of an artificial transcription factor (ATF) that induces endogenous interferon gamma expression. The transcription factor contains zinc finger proteins to recognize the interferon gamma gene and an activation domain to promote transcriptional activation. We report an active construct, ATF 2, that increased interferon gamma levels 14-fold in human T cells. Biophysical characterization of recombinantly expressed ATF2r provided information that zinc is required to prompt protein refolding. DNA binding assays show a KD of 5.27 + 0.3 nM. Thermal stability studies show an unfolding temperature of 52C for ATF2r. This work will provide a new and translational tool for regulated production of immune proteins to improve immunotherapy outcomes.
Rights
© 2024, Ashley King
Recommended Citation
King, A.(2024). Design of An Artificial Transcription Factor for Inducing Potent Interferon-γ Signals. (Master's thesis). Retrieved from https://scholarcommons.sc.edu/etd/7796