@unpublished{pittir38097,
           month = {January},
           title = {OPTICAL AND SMALL-MOLECULE CONTROL OF PROTEIN FUNCTION THROUGH GENETIC CODE EXPANSION},
          author = {Ji Luo},
            year = {2020},
        keywords = {genetic code expansion, unnatural amino acid, protein function},
             url = {http://d-scholarship-dev.library.pitt.edu/38097/},
        abstract = {Expanding the genetic code for site-specific incorporation of various biophysical probes and labels, such as fluorescent probes, photolabile caging groups, optical probes, photoswitches, bioorthogonal chemselective groups, offers a sophisticated tool for exploring protein structure and function, dissecting the cellular processes, and developing proteins with novel properties. Site-specific installation of distinct functional groups on a protein of interest is achieved using orthogonal aminoacyl-tRNA synthetase/tRNA pairs in response to an amber stop codon (UAG) placed in the gene of interest. In this dissertation, genetic encoding of a series of diverse unnatural amino acids (UAAs), including photocaged lysines, coumarin lysines, phthalimide lysines, photocaged cysteines, photocaged tyrosines, photoisomerizable phenylalanines, and azido lysines, were achieved through the discovery of the engineered pyrrolysyl-tRNA synthetase/tRNA pair from bacteria to eukaryotic cells. Strategic placement of these functional groups renders the protein inactive until deprotection through a bioorthogonal external trigger delivers the active wild-type protein. This developed methodology enables the conditional control of various cellular processes with spatiotemporal precision in a non-invasive way, including enzymatic activation, protein folding, protein translocation, gene repair (UvrD), post-translational modification (SUMO1), DNA recombination (Cre), RNA processing (Csy4), and gene editing (CRISPR/Cas9).}
}