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Showing 1 to 15 of 15 for “"Genetic code expansion"”.
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Genetic Code Expansion in Mammalian Cells
… scientists have developed methods to expand the genetic code of living organisms, introducing new, non-canonical amino acids with diverse chemistries into proteins. These methods rely on engineering of the translational machinery often importing an aminoacyl-tRNA synthetase (aaRS)/tRNA pair that …
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Site-Specific Labelling of the Flavivirus Envelope Protein using Genetic Code Expansion
… dynamic ZIKV particle. With this goal, I applied genetic code expansion to incorporate the non-canonical amino acid BCN-K at specific sites in the E protein. BCN-K is a versatile non-canonical amino acid that can be labelled via reactions with both tetrazine- and azide-conjugated probes. I …
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Identification and Evolution of New Orthogonal Aminoacyl-tRNA Synthetase/tRNA Pairs for Genetic Code Expansion
Genetic code expansion is the branch of molecular biology aiming to expand the repertoire of amino acids which can be incorporated into proteins in vivo. A central challenge in expanding the genetic code of cells to incorporate non-canonical amino acids is the scalable discovery of aminoacyl-tRNA …
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Synthesis, Characterisation and Evolution of Escherichia coli with a Compressed Genetic Code
… the creation of organisms with a compressed genetic code. Such liberated codons can be reassigned to non-canonical amino acids to equip proteins with novel functions. Furthermore, recoded organisms are genetically distinct from all other lifeforms and may therefore prove useful for …
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Mechanistic study and protein engineering of a dye-decolorizing peroxidase from Thermomonospora curvata for enhanced and expanded functions
… rational engineering, cofactor substitution, and genetic code expansion. In the first part of this work, we identified a key surface-exposed loop residue, Glu293, as a target for tuning substrate access and radical stability. Replacing E293 with glycine (E293G) enhanced peroxidase activity toward …
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Bioorthogonal Tethering of Drug Fragments to Engineered G Protein-Coupled Receptors
… This thesis presents a novel strategy employing genetic code expansion (GCE) and bioorthogonal chemical reactions for tethering drug fragments adjacent to allosteric sites in GPCRs to increase their potency and enable fragment-based drug screening in cellular systems. C-C chemokine receptor 5 …
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Expanding the genetic code of a mouse
Genetic code expansion has used the site-specific insertion of unnatural amino acids into proteins(Greiss and Chin, 2011). An aminoacyl-tRNA synthetase and a tRNA are used to specifically insert the unnatural amino acid during mRNA translation, in response to an amber stop codon (UAG) placed at a …
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Studies on approaches for sequencing cytosine modifications and the identification of their protein readers
Alongside the genetic code, epigenetic mechanisms including covalent cytosine modifications facilitate dynamic changes in gene expression throughout development and differentiation. Such modifications are also drivers and markers of disease, earning them great academic and clinical interest. A …
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Acetylation regulates Thioredoxin Reductase activity and oligomerization
… enzyme in the Trx system and contains the 21st genetically encoded amino acid, selenocysteine (Sec). There were multiple experimentally identified TrxR acetylation sites with an unknown effect on TrxR activity. My thesis tested the hypothesis that programmed protein acetylation will enhance the …
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SORTASE AND CHEMICAL BIOLOGY AS TOOLS TO STUDY UBIQUITINATION
… sequence in combination with click chemistry and genetic code expansion to develop a strategy for site-specific modification of proteins of interest with user-defined polyubiquitin chains. We exemplify the broad utility of this strategy for multiple different proteins of interest, including DNA …
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A General Platform for Ribosome Engineering
… Emerging research aims to reprogram the genetic code in vitro and in vivo to assemble non-canonical polymers with this approach. However, the properties of the natural ribosome limit existing work to close analogues of the canonical amino acids. This thesis develops a directed evolution …
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Using Translational Switching to Investigate the SCN Clockwork
… expression of a transgenic copy. By using genetic code expansion (GCE), it is possible to create translational switches whereby the translation of a transgene is conditional on the provision of a non-canonical amino acid. Incorporation of an ectopic amber stop codon (TAG) into the coding …
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Bioorthogonal Functionalization of Elastin-like Polypeptides
… of proteins will, by definition, require expansion of the biochemical toolbox; there are a variety of techniques used to achieve this goal. In these studies, we explore the use of genetic code expansion for incorporation of unnatural amino acids. This technology permits co-translational …