Research Overview: The Truex Lab develops protein technologies to study fundamental principles of protein folding and to create new therapeutic strategies for cancer immunotherapy.
Artificial Transcription Factors for Immunotherapy: This project seeks to develop transcription factor therapeutics that activate immune genes and direct their function. Most immunostimulatory agents act through extracellular receptors and signaling pathways, yet often face challenges when tumors and viruses suppress the signals required for immune recognition. Direct control of immune gene expression offers the promise of overcoming the limitations of current immunotherapies and enabling more precise antitumor responses.
To address these limitations, we develop artificial transcription factors that directly activate immune genes at the transcriptional level. We design, prepare, and study zinc-finger transcription factors that recognize defined DNA sequences and activate genes that mediate antitumor and antiviral immunity. Our current work focuses on transcription factors that activate interferon-γ expression and increase antigen expression. Students characterize DNA binding, transcriptional activity, immune function, and antitumor effects in biochemical and cellular assays and preclinical models. Training also includes methods to characterize immune cell function by immunoassays, flow cytometry, and transcriptomic profiling.
Engineered Fold-Switching Proteins for Expanding the Folding Code: This project seeks to define how small chemical interactions drive unusually large structural transitions in fold-switching proteins. These proteins are defined by their capacity to adopt an alternative stable fold, regulating biochemical processes that respond to subtle cues or mutations. How fold-switching proteins access multiple distinct states remains largely unknown across diverse biological systems, including KaiB in circadian regulation, XCL1 in immune defense, and RfaH in gene expression. Uncovering the rules of fold switching at the molecular level will unlock new protein designs that respond to chemical and biological signals.
We design, prepare, and study engineered fold-switching proteins to map precise molecular features onto structural and energetic changes between folds. We use automated flow protein synthesis and recombinant expression to prepare these proteins. To isolate the interactions that stabilize each conformation, we introduce sequence changes using natural and non-natural amino acids and covalent modifications. Students learn protein synthesis, expression, chromatographic purification, and mass spectrometry. Training also includes methods to characterize protein structure, stability, binding, and exchange by circular dichroism, isothermal titration calorimetry, and NMR spectroscopy.