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 17 of 17 for “"CRISPR-Cas9 screening"”.
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Identification of novel synthetic lethal interactions using multiplexed CRISPR-Cas9 screening
… interactions 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 …
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Defining cell state regulators in cancers using single-cell analysis and CRISPR-Cas9 screening
… screened using a single-cell RNA-seq coupled CRISPR- knock-out approach, with regulator potential uncovered for multiple targets including SOX10, MITF, EIF3G, PRPF19, RPS27A, and CDC20. Cell line annotations achieved through previous high-throughput screens were also leveraged to uncover …
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ESTABLISHING AN IN VITRO MODEL FOR CRISPR-CAS9 SCREENING OF INTERFERON EFFECTORS IN HBV INFECTION
… response and sought to identify them via a CRISPR-Cas9 knockout screen targeting ~2,000 ISGs. Identifying the ISGs that mediate antiviral activity may provide insights into how interferon responses can be potentiated, both to improve patient outcomes and to increase our understanding of …
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Application of CRISPR/Cas9 screening to study cancer drivers and to identify novel cancer vulnerabilities
… it simpler to accurately identify interactions. CRISPR/Cas9 technology was applied as it allows for large-scale, unbiased screening of genetic interactions. A genome-wide guide RNA library was prepared and implemented for knockout screening in the isogenic cell line panel. Analysis was performed …
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Characterisation of HIF2A targets identifies MYC as a key determinant of HIF2A dependency in ccRCC
… could then be functionally tested by *in vivo* CRISPR/Cas9 screening and subsequent rescue experiments. CRISPR/Cas9 screening revealed that *MYC* and *CCND1* are key dependencies in ccRCC *in vivo*, in addition to highlighting key differences between *in vitro* and *in vivo* dependencies. By …
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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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Utilizing in vivo genome-wide CRISPR/Cas9 screens in immunocompetent mouse models to identify novel tumor liabilities and explore mechanisms of resistance to Chimeric Antigen Receptor T-cell therapy
… (TME). Fortunately, with the advent of CRISPR/Cas9 screening platform technologies, mechanisms of treatment resistance and novel therapeutic target identification can be efficiently interrogated in an unbiased, high-throughput manner. Furthermore, adaptation of these platforms for use in …
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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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The identification of novel factors required for the development of primordial germ cells
… in vitro. Secondly, we coupled this system with CRISPR/Cas9 screening technology, which permitted us to identify novel factors required during the development of PGCLCs. We identified the translesion synthesis (TLS) factor Rev1 as a key factor involved in PGCLC development. Furthermore, we …
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Identification and functional characterisation of gene fusions in human cancer cell lines
… In this study, I combine RNA sequencing, CRISPR/Cas9 screens and high-throughput drug sensitivity data in a panel of 1,011 human cancer cell lines across 42 tissue types to examine the occurrence and functional relevance of gene fusions in cancer. Fusions were called using three algorithms …
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Applications of forward genetic screens to LncRNAs, cancer immunotherapy, and cellular engineering
… mapped to phenotypic changes. Recently, CRISPR-Cas9 has emerged as a powerful genetic perturbation technology, opening up new opportunities for forward genetic screens. In this thesis, I present work to advance this approach and demonstrate its application in a range of contexts relevant …
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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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Genetic alterations defining human primary melanoma and mechanisms of immune evasion
… intersect my genomic analysis with a dataset of CRISPR-Cas9 dropout screens and discover a melanoma-associated genetic vulnerability mediated by Interferon Regulatory Factor 4 (IRF4). I then begin to experimentally validate and explore the biological pathway by which IRF4 may function in the …
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Dissecting the early steps of MLL induced leukaemogenic transformation using a new mouse model of AML
… MLL-ENL expression. To this end, genome-wide CRISPR-Cas9 screening was performed on the new AML model developed. The data obtained were integrated with scRNA-seq results in order to identify and validate new potential therapeutic targets associated with early MLL-ENL driven transcriptional …
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Developing Targeted Therapies for T-cell Acute Lymphoblastic Leukemia
… drivers for dasatinib sensitivity. Genome-wide CRISPR/Cas9 screening, phosphorylation flow cytometry, and phosphorylation proteomics assays were employed to confirm the dependency and inhibitory effect of dasatinib on its therapeutic targets (dasatinib sensitivity drivers). In vitro …
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Viral- and host-targeted interventions to improve influenza disease outcomes
… during severe influenza infections. Using CRISPR/Cas9 screening techniques, and a type I IFN reporter system, we identified G-protein coupled estrogen receptor 1 (GPER1) as a candidate regulator. Our subsequent experiments demonstrated that blocking GPER1 function with a selective …
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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. …