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 28 for “"Homology directed repair"”.
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Developing a new efficient CRISPR-Cas9 base Homology Directed Repair oriented gene editing plasmid for Streptomyces
… gene editing plasmid that works based on the homology directed repair system in Streptomyces. The fundamental goal of the study is to characterise the phenotype of a deletion mutant in the gene cluster, SCO6357-53, by using the newly developed CRISPR-Cas9 plasmid. Previous study has shown that …
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Homology-Directed Repair of Dna Lesions Generated by the Mismatch Repair Dependent Processing of Cytotoxic Guanine Adducts
… thymine during DNA replication. The Mismatch repair (MMR) pathway rectifies base mismatches. However, MMR is unable to correct these S6-MeG:T or O6-MeG:T mispairs. In fact, upon recognition of the lesions, MMR initiates a futile repair cycle resulting in apoptosis. Thus, MMR is required for …
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A Genetic Test of a Model for Two Activities of Fushi Tarazu Protein in Drosophila Melanogaster
… for intragenic complementation. CRISPR-mediated homology directed repair (HDR) was used to introduce engineered ftz alleles into the ftz locus. Subsequently, four ftz engineered alleles were constructed in vectors for reintroduction by Recombinase-mediated cassette exchange (RMCE). Despite using …
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The circadian Control of DNA end Resection
… ends (Non-Homologous End-Joining, NHEJ) and the repair of the lesion using a homologue sequence (Homology-Directed Repair, HDR). The choice between these two pathways is crucial for cell fitness and is controlled by an exquisite regulatory system. In this thesis, we uncovered a novel role for the …
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Modulation of DNA repair pathway after CRISPR/Cas9 mediated Double Stranded Break
… Targeted modifications can be introduced while repairing double strand breaks (DSBs), induced by the CRISPR/Cas9 system. The DSB is repaired by either non-homologous end joining (NHEJ) or homologous recombination (HR), and the repair is commonly processed through NHEJ because it is the dominant …
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EXAMINING THE ROLE OF THE XAB2 PROTEIN IN HOMOLOGOUS RECOMBINATION
<p>DNA double strand break (DSB) repair is critical to maintain genomic integrity and cell viability. DSBs can occur during the course of cell cycle during replication or transcription, or by exogenous agents such as chemicals or ionizing radiation. For my thesis, I studied homologous recombination …
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Creating CRISPR-Cas9 genome edited iPSC lines to model a patient-specific mutation in mitochondrial disease
… associated donor template were used to induce homology directed repair (HDR) the genome of iPSC and knock-in the patient mutation. Guide RNAs were designed and tested for efficiency before electroporation into wild type iPSC. Clonal cell lines were made by low-density seeding and manual colony …
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Investigating the role of chromatin modifications in CRISPR/Cas9 gene editing
… of chromatin modifications on the outcome of DNA repair were detected, with similar efficiencies of homology-directed repair (HDR) and non-homologous end joining (NHEJ) on maternal and paternal alleles. Upon further characterisation we discovered that the majority of types of insertions and …
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Cystic fibrosis gene repair: correction of ΔF508 using ZFN and CRISPR/Cas9 guide RNA gene editing tools
… designed in this thesis can mediate efficient homology-directed repair (HDR) with appropriate donor repair plasmids to correct CF-causing mutations in a CF cell line. Firstly, the most common mutation, ΔF508, was corrected using a pair of existing ZFNs, which cleave in intron 9, and the donor …
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Novel Structures of RAD51 Reveal Mechanisms in DNA Damage Repair and Genomic Stability
… of genomic stability by promoting the repair of DNA double-strand breaks and the protection of DNA replication forks. RAD51 functions alongside the tumour suppressor protein BRCA2 to catalyse DNA strand-exchange reactions which form an integral part of Homology-Directed Repair. RAD51 …
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Optimizing nuclear transgene expression and genome editing for high-value compound production in the green microalga Chlamydomonas reinhardtii
… Moreover, the interaction between efficient homology directed repair and the length of homologous regions for this repair mechanism was shown. In this work, homology directed repair was improved for CRISPR/Cas9 mediated double strand breaks, developing a guideline of homology arm lengths. …
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A Cloning-Free Recyclable System for CRISPR-Cas9 Mediated Mutant and Reversion Construction in Candida albicans Clinical Isolates
… SAT1 nourseothricin resistance gene to serve as repair templates. PCR-amplification of these cassettes with primers containing sequences homologous to the flanking regions of the target gene allow for high-efficiency homology-directed repair. We first targeted ADE2 for deletion, as homozygous …
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Analysis of The RING Domain And BRCT Repeats of BRCA1
… is necessary for BRCA1's interaction with DNA repair proteins BACH1, ABRAXAS, and CtIP. To test the importance of these two domains we generated cell lines and mouse models with point mutations that either eliminate the E3 ligase activity of Brca1 while maintaining its interaction with Bard1 …
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53BP1/Shieldin Counteract DSB Resection Through Fill-In Synthesis
… The role of 53BP1 in double strand break (DSB) repair outside of these three well-studied contexts is less clear, though it has been proposed that 53BP1 acts as a master regulator of so-called "DSB repair pathway choice," promoting classical non-homologous end-joining (cNHEJ) at the expense of …
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Re-engineering of Dendrimer-Based Lipid Nanoparticles for Efficient and Precise HDR-Mediated Gene Editing
… has been made toward precise editing of DNA via Homology Directed Repair (HDR) that requires careful orchestration of complex steps. Rather, many reports of in vivo gene editing rely on an error-prone mechanism called Non-Homologous End Joining (NHEJ). While this pathway is effective for …
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In Vivo Genome Editing: Proof of Concept in Neonatal and Adult Mouse Liver
… of AAV. Gene targeting, the ability to make site-directed changes to the genome, has been a powerful tool for genetic discovery. Until recently, the low efficiency of targeting has rendered it unlikely to be of therapeutic value for most genetic diseases. Zinc finger nucleases (ZFNs) are …
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Investigating β- and γ-secretase function and processing of APP with genetic and pharmacological tools
… clustering, and outline the creation of homology directed repair templates for endogenous tagging of the catalytic subunit of γS (PSEN1), as well as BACE1, with a genetic construct allowing for fluorescent visualization and optogenetic manipulation. I then create and test several genetic …
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Endogenous Reporter Systems for High-Throughput Functional Screening in Clear Cell Renal Cell Carcinoma
… marker at the 3’ end of their respective locus. Homology-directed repair (HDR)- and non-homologous end joining (NHEJ)-based CRISPR-Cas9 knock-in strategies were utilized for the development of these systems that involve homology-dependent and minicircle-aided integration, respectively. These …
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CRISPR/CAS9 mediated gene editing for the improvement of beef and dairy cattle
… a 90 nucleotide single stranded-oligonucleotide repair template, which allows for homology directed repair to introduce the desired substitution at those loci. Cells from these cattle that are genetically improved by this technique may be used for somatic cell nuclear transfer and resulting …
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Generation of a CRISPR-Cas9 mediated knock-in reporter for the GRIA3 candidate gene for schizophrenia
… (CRISPR) - CRISPR associated protein 9 (Cas9) homology directed repair (HDR) approach so that a donor reporter vector containing the Gaussia secreted luciferase gene and linked green fluorescent protein (GFP) or neomycin resistance gene (Neo) gene would be integrated directly under the control …
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