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 20 of 40 for “"CRISPRi"”.
-
Investigating the role of sufT in Mycobacterium abscessus using CRISPRi
… interspaced palindromic repeats interference (CRISPRi) system was recently optimized for transcription repression in mycobacteria. This study aimed to investigate the functionality of the mycobacterial CRISPRi in M. abscessus, specifically to investigate the role of the sufT homologue, …
-
Investigating the chromatin dynamics that underlie CRISPRi-validated enhancer-promoter interactions
… remained costly until recent developments of CRISPRi methods enabled large scale perturbation experiments for identifying the target genes of enhancers. However, CRISPRi methods produced results that were not always consistent with the previous understanding of enhancer-promoter interactions …
-
CRISPRI-based high-throughput functional genomic approaches for use in mycobacteria
… and nextgeneration sequencing, and is termed CRISPRi-Seq. A pooled library of 11 367 mutants, targeting 2 385 M. smegmatis genes with M. tuberculosis homologues, was constructed and used to infer gene essentialities which were compared with corresponding predictions from transposon-insertion …
-
CRISPRi screens to identify combination therapies for the improved treatment of ovarian cancer
… screens combining cisplatin or oxaliplatin with CRISPRi pooled gene knockdowns. Screens were analyzed at 9-days to mimic patient care, and at 48-hours to isolate the short-term DNA damage response. Genes whose knockdown caused sensitivity to the platinum chemotherapeutics were identified through …
-
Uncovering Functional Alzheimer’s Disease Variants and Their Effector Genes through Single-Cell CRISPRi Screening and 3D Genomics
… genes. Second, we designed a single-cell CRISPRi screen to perturb 74 candidate regulatory regions and assessed their transcriptional response in the human microglial cell line, HMC3. We collected 96,639 single cells and identified 19 regulatory regions with 19 corresponding effector …
-
Development and validation of a CRISPR interference system for gene regulation in Campylobacter jejuni
… a CRISPR-based interference system known as CRISPRi in C. jejuni to control gene expression and thereby investigate gene function. To achieve this, a “dead” cas9 and sgRNA backbone from the Streptococcus pyogenes CRISPRi system was combined with C. jejuni-derived promoters of predetermined …
-
Investigating the Role of RNA-Binding Proteins in Tumor Response to DNA Damage-Inducing Chemotherapy
… carcinoma (HGSOC) we performed a targeted CRISPRi screen in vitro against 579 RBPs to identify those that cause sensitivity or resistance to cisplatin, oxaliplatin, paclitaxel, or olaparib. The screen implicated several known and novel RBPs in affecting sensitivity to chemotherapy in HGSOC, …
-
A Chemical-Genetic Map of Drug Resistance in Mycobacterium Tuberculosis
… in Mtb using a novel CRISPR interference (CRISPRi)-based chemical genetics platform. Our lab developed a CRISPRi library containing 100,000 unique knockdown strains and treated this library with a panel of some of the most important antitubercular drugs. The fitness of each mutant in the …
-
Dynamic regulation of pathways by down-regulating competing enzymes
… laborious in Pseudomonas. Here, we developed a CRISPRi gene repression system by engineering the Streptococcus pasteurianus dCas9 and sgRNA. We demonstrate a robust and titratable gene depletion system, with up to 100-fold repression in [beta]-galactosidase activity in P. aeruginosa. We …
-
Identification and characterisation of HNF1B as a pro-tumorigenic lineage factor in clear cell renal cell carcinoma
… demonstrated that both HNF1B CRISPR knockout and CRISPRi knockdown conferred a strong selective disadvantage in vitro that was rescued by HNF1B restoration, suggesting that it was a specific effect of HNF1B loss. I also showed that HNF1B knockout impaired tumour initiation and growth in vivo. I …
-
Development of Improved Genetic Methods for Enterococcus Faecium
… and the roadmap to generating functional CRISPRi in vancomycin-resistant E. faecium (VREfm). By utilizing my integration system, I was able to create genome integrated inducible dCas9 controlled by IPTG or agmatine inducible systems in VREfm. Furthermore, I generate and characterize a …
-
Targeted depletion of RibF, a putative bifunctional FAD synthetase/ flavokinase in Mycobacterium smegmatis using CRISPR interference
… of downstream genes. CRISPR interference (CRISPRi) technology was used to generate an anhydrotetracycline (ATc)-inducible ribF hypomorph of M. smegmatis, a widely exploited mycobacterial model. Consistent with other organisms, ribF was shown to be essential for in vitro growth of M. …
-
Multiplexing cell-cell communication
… sub-circuits constructed from a total of 12 CRISPRi-based transcriptional logic gates, an acyl homoserine lactone-based communication module, and three inducible promoters that enable small molecule control over the conversations. Experimentally-parameterized mathematical models of the …
-
Hypoxia signalling regulates human lung epithelial cell fates
… hypoxia-induced airway differentiation. With CRISPRi and ectopic expression systems, I demonstrated that HIF-1 and HIF-2 differentially regulated airway and alveolar fate decisions of lung epithelial progenitors. Combining transcriptomic and targeted DamID-sequencing analyses, my collaborators …
-
Uncovering the biogenesis pathways for human mitochondrial alpha-helical outer membrane proteins using genome-wide approaches
… MTCH2 was discovered using a genome-wide CRISPRi screen for alpha-helical tailanchored substrates (Guna et. al, 2022), the broader biogenesis and targeting pathways for all biophysically diverse alpha-helical proteins remained unexplored. Critically, the mechanisms of cytosolic chaperoning …
-
Creating effective drug discovery methods in Mycobacterium abscessus
… a phenotypic screening selection funnel, a CRISPRi arrayed library, and transcriptomic predictions of drug action. Increasing the focus of antibiotic discovery has been on narrow spectrum, species specific compounds, and I have focussed on developing tools in a single bacterium, …
-
Probing the Mycobacterial Cell Envelope: CRISPR interference-mediated investigation of two essential genes in the galactan biosynthetic pathway
… and cell morphology. The CRISPR interference (CRISPRi) genome editing system was used to target genes involved in galactan biosynthesis and generate transcriptional knock down of wecA (also known as rfe, MSMEG_4947) and rfbD (MSMEG_6369) in M. smegmatis, a widely exploited modal for …
-
FtsK Regulates the Assembly of the Division Machinery in the FtsZ-less Chlamydia trachomatis
… with XerD during cell division. In addition, CRISPRi which uses a crRNA to target an inducible and defective Cas12 enzyme (dCas12) to a specific chromosomal sequence, has shown that chlamydial FtsK also colocalizes with a chromosomal sequence immediately adjacent to the dif site during cell …
-
Highly Saturated Transposon Sequencing identifies genes impacting Staphylococcus aureus pathogenesis in macrophages
… infection. We used CRISPR interference (CRISPRi) to functionally validate phenotypic contributions for a select subset of genes. Of the 20 genes passing validation, 7 had a previously identified role in S. aureus virulence, and 13 were newly implicated. Validated genes frequently …
Page 1 of 2