Tschann, Madison2021-02-222021-02-222018-12https://hdl.handle.net/11299/218674University of Minnesota M.S. thesis. December 2018. Major: Stem Cell Biology. Advisor: Susan Keirstead. 1 computer file (PDF); vi, 43 pages.Regulatory T-cells (Tregs) are a subset of T-cells essential for maintaining immune tolerance and their dysregulation has been found to have a central role in the progression of various autoimmune diseases. The transplantations of Treg as a form of immune therapy has and continues to be an attractive method for the treatment of such disease based on their immuno-modulatory properties. Despite its potential, Treg adoptive cell transfer therapy is hampered by limited isolation efficiency due to low frequencies in human peripheral blood and poor in vitro expansion of a pure population. Herein, a novel CRISPR/Cas9 based technique is described utilizing AAV incorporation of strong transcriptional elements into the promoter region of the Treg master transcription factor, FOXP3, to upregulate expression in isolated primary T-cells and drive them toward a Treg phenotype.enCRISPR/Cas9Genome editingGenome engineeringRegulatory T-cellsThesis or Dissertation