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 20 of 622 for “"Cas9"”.
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Cas9-induced on-target genomic damage
CRISPR/Cas9 is the gene editing tool of choice in basic research and poised to become one in clinical context. However, current studies on the topic suffer from a number of shortcomings. Mutagenesis is often assessed using bulk methods, which means rare events go undetected, unresolved or are …
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Epigenetic Determinants of CRISPR-Cas9 genotoxicity
… editing therapies, such as those using CRISPR-Cas9, offer promising treatment options for genetic disorders. However, they also present unique safety assessment challenges including detection of off-target effects and unintended genomic alterations. An unresearched area of these concerns …
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Modulation of the CRISPR-Cas9 Immune Response
… spacers. In type II-A CRISPR systems, Cas9 uses the guide RNA to cleave target DNA sequences in the genome of infecting phages, and the tracrRNA to bind the promoter of cas genes and repress their transcription. We previously isolated a Cas9 mutant carrying the I473F substitution that …
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Constructing and deconstructing cancer using CRISPR-Cas9
… part of this thesis, I describe a novel CRISPR-Cas9-based approach for rapid functional investigation of candidate genes in vivo using well established autochthonous mouse models of cancer. By employing this platform in a GEMM of lung adenocarcinoma in vivo, I have functionally validated both …
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Cas9-Primed Adaptive Immunity During the CRISPR-Cas Response
… CRISPR-Cas systems, spacer sequences direct the Cas9 nuclease to target infecting bacteriophages and cleave their double-stranded (ds)DNA genomes. Whether and how pre-exiting anti-viral spacers in type II systems affect memory generation and the acquisition of new spacers is unknown. Here, in my …
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Using CRISPR/Cas9 to modify the genome of cattle
… expanded the possibilities of gene editing. The Cas9 nuclease creates a double strand break which can be repaired by non-homologous end joining (NHEJ) or homology directed repair (HDR). The goal of this experiment was to create a single nucleotide change using the CRISPR/Cas9 system in …
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Focused CRISPR-Cas9 screens investigating the DNA damage response
… opportunities. We demonstrate that CRISPR-Cas9 gene editing is a useful tool in genetic screens investigating the DDR. We present the results of three screens using focused CRISPR-Cas9 guide RNA libraries each interrogating different aspects of DDR biology. Firstly, we used a kinase-focused …
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Harnessing CRISPR-Cas9 screens to identify functional genetic interactions
The advent of CRISPR-Cas9 genetic engineering technologies has vastly improved our ability to interrogate genetic interactions in high-throughput studies. CRISPR-Cas9 screens have rapidly evolved to become the main approach for unbiased, genome-wide, forward genetic studies. This work seeks to …
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Genetic screens in vivo using the CRISPR/Cas9 system
An unmet and paramount need in the field of cancer research is to rapidly translate basic biological findings to clinically relevant therapeutics for cancer patients. Recent technological advances have generated many innovative applications to cancer biology and in a short time have yielded a …
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A novel CRISPR-Cas9 platform with divergent targeting capabilities
… highly-similar, yet previously uncharacterized, Cas9 from Streptococcus canis (ScCas9) through rational manipulation of distinguishing motif insertions. To this end, we report a divergent affinity to 5'-NNGT-3' PAM sequences, as well as preferences for expanded 5'-NNG- 3' motifs, and demonstrate …
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CRISPR/Cas9-mediated ATG7 knockout results in enhanced osteosarcoma chemosensitivity
… chemotherapy drugs. In the current study, CRISPR/Cas9 was used to knockout (KO) ATG7, a key gene in the autophagy process, in HOS-143B (highly metastatic/aggressive) osteosarcoma cells. Successful knockout of ATG7, and the autophagy process, was validated by qPCR and cell viability assays. …
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The Interrogation of Cas9 Aptamers and sgRNA Structures Through SELEX
… much of the current focus on advancing CRISPR-Cas9 editing revolves around the engineering of Cas9, the interrogation and evolution of sgRNA scaffold, in addition to novel Cas9 binding RNAs, represent another echelon of development and therapeutic potential. Currently, the majority of research …
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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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Development of CRISPR/Cas9-based gene drive biotechnology in S. cerevisiae
… cerevisiae as a model to develop CRISPR/Cas9-based gene drive biotechnology. The CRISPR/Cas9 biotechnology is a powerful gene editing tool used to modify target genomic sequences in all forms of living creatures including animals, plants, fungi, and bacteria. One potential application of …
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Biases and Blind-Spots In Genome-Wide Crispr-Cas9 Knockout Screens
<p>Adaptation of the bacterial CRISPR-Cas9 system to mammalian cells revolutionized the field of functional genomics, enabling genome-scale genetic perturbations to study essential genes, whose loss of function results in a severe fitness defect. There are two types of essential genes in a cell. …
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Generation of Memory of Infection During the CRISPR-Cas9 Immune Response
… into small RNA molecules that guide the Cas9 nuclease to the viral targets (protospacers). Streptococcus pyogenes Cas9 cleavage of the viral genome requires the presence of a 5′-NGG-3′ protospacer adjacent motif (PAM) sequence immediately downstream of the viral target. Before my graduate …
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Investigating the role of chromatin modifications in CRISPR/Cas9 gene editing
… heterochromatin have been shown to impede CRISPR/Cas9 editing efficiency. Conversely, Cas9 can open previously inaccessible regions of DNA, and transcriptionally silent targets can usually be edited without difficulty. In order to address this paradox, we have developed a method that exploits the …
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Identification of novel synthetic lethal interactions using multiplexed CRISPR-Cas9 screening
… and built a bespoke multiplex CRISPR-Cas9 library to interrogate 1192 gene-pair interactions. We deployed this library on a panel of melanoma cell lines and a retinal pigment epithelial cell line as a normal comparator. We went on to develop a bioinformatic pipeline with which to …
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