Global ETD Search
Search theses and dissertations gathered from participating repositories worldwide. Every result links back to the library that holds it. No account is needed.
Results
Showing 1 to 19 of 19 for “"Base editing"”.
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Base editing of Galactose-1-Phosphate Uridylyl Transferase as a novel gene therapy approach to treat Q188R mutation in a cellular model of Classic Galactosemia
… galactitol. The Q188R mutation is caused by a base change from adenine to guanine in base pair 563. This mutation causes an amino acid change in position 188 from glutamine (Gln) to arginine (Arg) and is responsible for 60-70% of CG cases in homozygous state. Consequently, affected infants …
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Applications of mitochondrial gene therapy
… defect prior to the advent of mitochondrial base editing. In particular, the combined use of base editing and nuclease treatments showed greater reduction in mutation burden as well as a reduction in the off-target effects associated with base editing. In summary, this work expanded the …
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Modeling structured biological processes with machine learning
… model fundamental biological processes in genome editing and directed evolution. I first consider a model of DNA repair following CRISPR/Cas9 cleavage, which was generally thought to be unpredictable. In a large-scale dataset, I find signatures implicating an alternative and more predictable DNA …
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CRISPR gene editing for Cystic Fibrosis: targeting the G542X mutation with base editor engineered virus-like particles
… care and face a reduced life expectancy. CRISPR-based gene editing offers the potential for a cure, by addressing the underlying cause of the disease. The doctoral work presented in this dissertation explored the avenue of CRISPR therapeutic gene editing for CF, while also testing and …
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Precision Gene Editing for Muscle Diseases
… effect on the afflicted individual. Gene editing tools like base editors and prime editors enable us to target and correct the underlying genetic causes of these diseases. Base editors enable precise base pair transition edits, while prime editors offer even more versatility, allowing for …
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Genetic Engineering as a Tool to Investigate Mitochondrial Gene Expression
… mitochondria, in particular the development of base editing, it is now possible to investigate these processes in more detail than ever before. Using these reverse genetics approaches, this work aims to utilise this new toolkit to probe different aspects of mitochondrial gene expression and …
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Quantitative modelling of CRISPR-Cas editing outcomes
… In this thesis, I recount the history of gene editing from transgenesis to prime editors. I then review the modelling techniques I use in my projects, before covering the state of predictive modelling for genome engineering. In two main results chapters, I discuss my work on modelling base …
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Determining the Roles of Q-SNAREs in Mast Cell Tumor Necrosis Factor Exocytosis
… -4, -6, and –11 KO RBL-2H3 cells using Cytosine Base Editing, and successfully generated heterozygous KO cell lines of all four genes. The results of these investigations have revealed that Stx2 may be dispensable for mast cell exocytosis in general potentially significant roles for Stx3 and …
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Discovery of novel CRISPR enzymes for transcriptome engineering and human health
… specific, across many transcripts tested. RNA editing with Cas 13 was also highly efficient, with up to 90% base editing rates, and as low as 20 off-targets with engineered specificity versions. Lastly, we combined Cas13 with isothermal amplification to develop a CRISPR-based diagnostic …
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Leveraging cellular models of polycystic kidney disease for mechanistic discovery and therapeutic development
… In our kidney organoid model, we utilize CRISPR base editing to generate human pluripotent stem cells with specific clinically documented patient mutations. This system elicits a robust cystic phenotype and allows evaluation of therapeutic interventions. We show that partial restoration of …
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Development of biophysical and Mass Spectrometry Assays for APOBEC3 cytosine deaminases: discovery and validation of cnvalent and non-covalent ligands
… that catalyze the conversion of a cytosine base to uracil in single-stranded DNA. There are seven members of the APOBEC3 family – A3A/B/C/DE/F/G/H. APOBEC3 proteins are a part of the innate immune defense system against viral infection, and mutate the viral genome to restrict replication. …
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Exogenous donor-derived hepatogenesis in chimeric hosts
… a generalized introduction to advanced genome editing techniques was provided. In Chapter II, we outlined optimized strategies for generating Hhex and Fah knockout embryos using CRISPR/Cas9 and base editing techniques. The outlined methods tested the latest genetic engineering techniques and …
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CRISPR AS A TOOL FOR TARGET DISCOVERY AND TECHNOLOGICAL INNOVATION: HIGH-THROUGHPUT CBE SCREENING TO EVALUATE M6A MODIFICATION PATHWAY IN CANCER PROLIFERATION AND DRUG RESISTANCE, AND INNOVATIVE PRIME EDITING STRATEGIES FOR DISEASE MODELLING AND THERAPY
… numerous effectors, which are classified based on their function. m6A is deposited co-transcriptionally by “writers”, including methyltransferase-like 3 (METTL3) and methyltransferase-like 14 (METTL14) that form a heterodimer regulated by the METTL-associated complex (MACOM). Its removal …
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A study of the causes and consequences of SFTPC mistrafficking in alveolar type 2 cells using novel organoid models
… expression and its endogenous expression using base editing. These heterozygous models reproduced key phenotypes observed in patient tissue and demonstrated numerous defective cellular pathways including altered apicobasal polarity, perturbed lumenogenesis, global endolysosomal dysfunction, and …
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Determining therapeutically actionable genetic interactions in human cancer at scale using multiplexed CRISPR screening
… identify SL interactions. A novel tRNA-based dual guide expression system was used to construct the Synergy library, enabling comprehensive and unbiased monogenic and digenic perturbation of 931 prioritised gene pairs. These pairs were identified through two large-scale bioinformatic …
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Engineering TEV Protease Specificity: An Exploration of Machine Learning and High-Throughput Experimentation for Protein Design
… However, many previous experimental and physics-based attempts at protease engineering have failed to engineer specificity in cleaving alternative substrates, rendering them useless. In this thesis, we aim to engineer TEV (tobacco etch virus) protease, a highly sequence-specific protease, to …
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Inferring effective cancer combination therapies using network-based multi-omics data integration
… suppression, causing resistance to monotherapy-based interventions. Drug combination therapies hold the promise of higher efficacy by simultaneously targeting compensatory mechanisms in tumours and reducing toxicity through the use of lower drug doses. However, identifying effective combination …
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Genomic Characterization of Sickle Cell Mouse Models for Therapeutic Genome Editing Applications
… we showed that CRISPR-Cas9-mediated genome editing can recreate a naturally occurring HPFH variant in the γ-globin (HBG1 and HBG2) promoters. Disruption of a TGACC nucleotide motif within this region by Cas9-mediated non-homologous end joining in human erythroid cells or their progenitors …
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Investigation of Clinically Relevant Fluconazole Resistance Mechanisms in the Fungal Pathogen Candida parapsilosis
… CpCDR1B and CpCDR1C on triazole MICs. The single base editing system was also used to place the SNP leading to the Y132F substitution into the triazole drug target CpErg11 of susceptible isolates. Antifungal susceptibility testing demonstrated this frequently cited driver of resistance was …