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Development of CRISPR-Cas systems for genome and transcriptome editing in fusarium circinatum
Dissertation (MSc (Microbiology))--University of Pretoria, 2024.
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| Format: | Thesis |
| Language: | English |
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University of Pretoria
2024
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| _version_ | 1867613469129310209 |
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| access_status_str | Open Access |
| author2 | Steenkamp, Emma Theodora |
| author_browse | Steenkamp, Emma Theodora |
| author_facet | Steenkamp, Emma Theodora |
| collection | Thesis |
| dc_rights_str_mv | © 2023 University of Pretoria. All rights reserved. The copyright in this work vests in the University of Pretoria. No part of this work may be reproduced or transmitted in any form or by any means, without the prior written permission of the University of Pretoria. |
| description | Dissertation (MSc (Microbiology))--University of Pretoria, 2024. |
| format | Thesis |
| id | oai:repository.up.ac.za:2263/94992 |
| institution | University of Pretoria (South Africa) |
| language | English |
| last_indexed | 2026-06-10T12:36:38.421Z |
| license_str | Other — see source repository |
| provenance_str_mv | Harvested via OAI-PMH from UPSpace — University of Pretoria Institutional Repository |
| publishDate | 2024 |
| publishDateRange | 2024 |
| publishDateSort | 2024 |
| publisher | University of Pretoria |
| publisherStr | University of Pretoria |
| record_format | dspace |
| source_str | UPSpace — University of Pretoria Institutional Repository |
| spelling | oai:repository.up.ac.za:2263/94992 Development of CRISPR-Cas systems for genome and transcriptome editing in fusarium circinatum Steenkamp, Emma Theodora alida.vandijk@up.ac.za Wingfield, Brenda D. Van Dijk, Alida UCTD Pathogenicity Growth and sporulation gene Fusarium circinatum CRISPR-Cas9 genome editing Virulence CRISPR-Cas13 transcript editing Sustainable Development Goals (SDGs) SDG-09: Industry, innovation and infrastructure Natural and agricultural sciences theses SDG-09 SDG-15: Life on land Natural and agricultural sciences theses SDG-15 Dissertation (MSc (Microbiology))--University of Pretoria, 2024. Fusarium circinatum, the causal agent of pine pitch canker disease, poses a significant threat to the health of Pinus species worldwide. Infection by this pathogen results in resinous canker formation in adult pine trees, and root and collar rot in seedlings, resulting in a severe mortality rate. While much is known about this pathogen, there is a lack in the understanding of the underlying virulence mechanisms. To date very few genes have been functionally characterized and shown to play a role in F. circinatum pathogenicity which is likely due a lack of efficient genome and transcriptome editing systems. For this purpose, we established a CRISPR-Cas9 and CRISPR-Cas13 system to enable editing at the DNA and RNA levels, respectively. The CRISPR-Cas9-mediated gene knockout system was established in F. circinatum to functionally characterize a putative pathogenicity gene. The pgs gene was found to play a role in pathogenicity, growth and sporulation when comparing various knockout isolates to their respective wild type isolates in various phenotypic assays. The CRISPR-Cas13 system was developed and established in F. circinatum as a proof of concept for the in vivo use of the RNA-targeting system in filamentous fungi. The system was capable of targeting GFP transcripts in GFP-expressing isolates resulting in a decrease in GFP fluorescence. This research expands the molecular toolbox available for functional characterization, RNA regulation, and editing, not only in F. circinatum, but in related species as well. National Research Foundation (NRF) through a DST/NRF SARChI chair in Fungal Genomics MSc bursary (Grant number: 98353). Microbiology and Plant Pathology MSc (Microbiology) Restricted Faculty of Natural and Agricultural Sciences 2024-02-29T08:32:35Z 2024-02-29T08:32:35Z 2024-04-16 2024-02-28 Dissertation * A2024 http://hdl.handle.net/2263/94992 https://doi.org/10.25403/UPresearchdata.25304680 en © 2023 University of Pretoria. All rights reserved. The copyright in this work vests in the University of Pretoria. No part of this work may be reproduced or transmitted in any form or by any means, without the prior written permission of the University of Pretoria. application/pdf University of Pretoria |
| spellingShingle | UCTD Pathogenicity Growth and sporulation gene Fusarium circinatum CRISPR-Cas9 genome editing Virulence CRISPR-Cas13 transcript editing Sustainable Development Goals (SDGs) SDG-09: Industry, innovation and infrastructure Natural and agricultural sciences theses SDG-09 SDG-15: Life on land Natural and agricultural sciences theses SDG-15 Development of CRISPR-Cas systems for genome and transcriptome editing in fusarium circinatum |
| title | Development of CRISPR-Cas systems for genome and transcriptome editing in fusarium circinatum |
| title_full | Development of CRISPR-Cas systems for genome and transcriptome editing in fusarium circinatum |
| title_fullStr | Development of CRISPR-Cas systems for genome and transcriptome editing in fusarium circinatum |
| title_full_unstemmed | Development of CRISPR-Cas systems for genome and transcriptome editing in fusarium circinatum |
| title_short | Development of CRISPR-Cas systems for genome and transcriptome editing in fusarium circinatum |
| title_sort | development of crispr cas systems for genome and transcriptome editing in fusarium circinatum |
| topic | UCTD Pathogenicity Growth and sporulation gene Fusarium circinatum CRISPR-Cas9 genome editing Virulence CRISPR-Cas13 transcript editing Sustainable Development Goals (SDGs) SDG-09: Industry, innovation and infrastructure Natural and agricultural sciences theses SDG-09 SDG-15: Life on land Natural and agricultural sciences theses SDG-15 |
| url | http://hdl.handle.net/2263/94992 https://doi.org/10.25403/UPresearchdata.25304680 |