Study Of Intracellular Sugar Driven Effects Utilizing Peptides, Proximity Labeling, And Crispr Based Techniques
Abstract
O-linked N-acetylglucosamine (O-GlcNAc) is a vital post-translational modification that impacts over 8,000 mammalian proteins at over 7000 serine and threonine residues. One of the proteins impacted by this modification is Sox2, a key regulator of cell stemmness and pluripotency that is often misregulated in cancers. Study of O-GlcNAc driven effects in this work utilizing proximity labeling as it relates to the transcription factor Sox2 has revealed a differential set of proteins that is enriched up on High or Low O-GlcNAc states. Additional work has also been done to expand this a set of known O-GlcNAcylated proteins utilizing CRISPR based knock-in techniques to allow for the installation of GFP, NanoLuc, and HaloTag proteins on the C-terminus of several crucially O-GlcNAcylated proteins in mammalian cells.