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.
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Showing 1 to 19 of 19 for “"CRISPR/Cas9 screen"”.
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Genomewide Crispr/Cas9 Screen Identifies Network of Protein Complexes That Regulate Trim24
… this possibility, I performed a genomewide CRISPR/Cas9 screen library using fluorescence activated cell sorting (FACS) to identify regulators of TRIM24. The screen was enabled by two innovations. I engineered cells with an in-frame knock-in of mClover3 to the endogenous copy of TRIM24 to …
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Identification of Novel Regulatory Genes in Acetaminophen Induced Hepatocyte Toxicity by a Genome-Wide CRISPR/Cas9 Screen
… or liver injury. This study used a genome-wide CRISPR/Cas9 screen to evaluate genes that are protective against or cause susceptibility to APAP-induced liver injury. HuH7 human hepatocellular carcinoma cells containing CRISPR/Cas9 gene knockouts were treated with 15mM APAP for 30 minutes to 4 …
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Identification of host factors required for cytosol entry by Shigella flexneri: Results of a genome-wide CRISPR/Cas9 screen
… I developed and carried out a genome-wide CRISPR/Cas9 knockout screen using a tissue culture model of epithelial cell infection. I used a hierarchical validation method which combined arrayed high-content microscopy and semi-automated image analysis to confirm a novel set of host factors …
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Focused CRISPR-Cas9 screens investigating the DNA damage response
… syndromes to developmental disorders. Genetic screens can be used to gain insights into the function of known DDR proteins, identify novel DDR components and highlight potential therapeutic opportunities. We demonstrate that CRISPR-Cas9 gene editing is a useful tool in genetic screens …
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SMARCB1 Maintains Lineage Fidelity in Clear Cell Renal Cell Carcinoma
… lineage factor dependence in cancer. Using CRISPR/Cas9 loss-of-function screening coupled with in vitro and in vivo validation I show that the loss of SMARCB1, a member of the SWI/SNF chromatin remodelling complex, confers an advantage to ccRCC cells upon inhibition of the essential renal …
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Investigating the regulation of DNA non-homologous end-joining through Ku70/80 interacting factors
… WRN knockout cells were generated and a CRISPR-Cas9 screen performed to identify suppressors of WRN sensitivity to DNA damage. The targets identified offer insights into WRN function.
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Inferring context-specific essentiality networks using large-scale CRISPR-KO screens
Large-scale genome-wide CRISPR knockout screens, such as the ones from DepMap and Project Score, revealed that a lot of genes are essential, i.e. required, in only a subset of cell lines. These context-essential genes offer insights into vulnerabilities of different cancer types and provide …
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The role of the histone variant H3.3 and its chaperones in the response to DNA damage
… cell system, the lymphoblastoid line TK6. Using CRISPR-Cas9 genome editing, I created H3.3, ATRX and HIRA knockout mutants and investigated their response to UV irradiation. h3.3 TK6 cells are not hypersensitive to acute DNA damage but exhibit persistence of DNA damage markers. h3.3 cells also …
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Interrogating DNA replication stress and DNA damage responses via phenotypic and functional CRISPR/Cas9 screens
… hypersensitivity, we performed a genome-wide CRISPR/Cas9 screen in ATM-deficient cells treated with the topoisomerase I poison topotecan. This dissertation describes two distinct resistance mechanisms in Atm-/- cells towards TOP1 inhibitors, resulting from the inactivation of the …
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Epitranscriptional Control of the Caspase-2—MDM2—p53 Axis: The Role of the m6A Writer in p53 Signaling.
… centrosome-derived stress. Using a genome-wide CRISPR/Cas9 screen coupled with a fluorescent reporter system, we uncovered all seven components of the m6A writer complex (METTL3, METTL14, WTAP, VIRMA, RBM15, ZC3H13, and CBLL1) as top-ranking hits required for proper activation of the …
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The role of a pore-forming protein, Mpeg1, in cytosolic import
… cytosol. Using the saporin assay, we performed a CRISPR/Cas9 screen targeting genes enriched in the dendritic cell (DC) subset most efficient at cytosolic import. With this approach we identified Mpeg1, as a potential player in cytosolic import in DCs. Mpeg1 is a member of the membrane attack …
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Genome-wide search for regulators of the first cell lineage decision in the mouse: a CRISPR/Cas9 endeavour
… to trophoblast in an unbiased approach, using CRISPR/Cas9 genome-wide loss of function screening. I developed a reporter cell line that constitutively expressed Cas9 from the Rosa26 locus, and was engineered with knock-in of T2A-Venus into the Elf5 locus (Elf5::Venus ESCs). Elf5 is a stringent …
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Exploring the role of endogenous DNA damage in neurodegeneration
… drugs. In addition, I performed a whole genome CRISPR/Cas9 screen. Subject to validation, the screen demonstrated that inhibition of proteins in key pathways, such as chromatin regulation and transcription, rescued the synthetic lethality between *Fam193a* and 5-fluorouracil. Second, I assessed …
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Defining Determinants of Primary Drug Resistance in Precision Cancer Therapies
… unbiased pharmacological and functional genetic screening approaches to overcome the persistent problem of primary drug resistance in two cancer contexts: (1) epidermal growth factor receptor (EGFR)-driven triple-negative breast cancer (TNBC) and (2) PIK3CA mutant gastric cancer. Particularly, in …
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The role of IFN-γ in cell-autonomous immune responses against Salmonella Typhimurium and Toxoplasma gondii
… described in Chapter 2 used a genome-wide CRISPR-Cas9 screen as an unbiased approach to identify new IFN-γ-induced genes that play a role in the cell death pathway. This viability screen enriched for cells resistant to LPS-mediated cell death after IFN-γ priming. Their sequencing generated …
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Targeting DNA Repair in Prostate Cancer: Therapeutic Combinations & Disease Models
… DNA damage response signalling. A whole genome CRISPR-Cas9 screen was performed to identify candidate genes that may drive sensitivity or resistance to combination treatment strategies. The findings from this work provide insights into the potential for targeting DNA repair in prostate cancer …
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OPTIMISATION AND THERAPEUTIC-TARGET-DISCOVERY ORIENTED ANALYSIS OF CRISPR-CAS9 SCREENS
The emergence of CRISPR-Cas9 technology has revolutionized cancer research by enabling targeted genome modifications in tumour cells, offering insights into cancer biology and potential treatment targets. My PhD research focuses on developing tailored tools and pipelines to enhance the analysis of …
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DISSECTION OF HOST-VIRUS INTERACTIONS IN THE HUMAN CENTRAL NERVOUS SYSTEM
… either IAV or MPXV. By developing a genome-wide CRISPR/Cas9 screening platform in human neurons, we were able to identify several candidate host factors that regulate neuronal anti-HSV-1 immunity, such as MTBP, SIRT7, GNAI3, NPPA, POLR3E and RNGTT. In a complementary approach, we performed high …
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Genome scale identification of HIV Dependency factors across multiple HIV-1 strains
In this thesis, I developed a method called HIV-CRISPR screening at genome-wide scale, and then at a smaller, targeted scale, to uncover novel genes that may act as HIV-1 dependency factors. By using a screening technique which assesses enrichment or depletion of guide based on HIV-1 release from …