DNA-free genome editing of Solanum tuberosum. A CRISPR/Cas9-mediated proteolistic approach for novel crop production

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dc.contributor.advisor Doyle Prestwich, Barbara en
dc.contributor.advisor Henriques, Rossana en
dc.contributor.author Meehan, Dara Eoin
dc.date.accessioned 2020-05-13T08:44:21Z
dc.date.available 2020-05-13T08:44:21Z
dc.date.issued 2020-05-10
dc.date.submitted 2020-05-10
dc.identifier.citation Meehan, D. E. 2020. DNA-free genome editing of Solanum tuberosum. A CRISPR/Cas9-mediated proteolistic approach for novel crop production. MRes Thesis, University College Cork. en
dc.identifier.endpage 49 en
dc.identifier.uri http://hdl.handle.net/10468/9936
dc.description.abstract Climate change is an ever-growing concern for global food security and crop production. Further adding to these issues is population growth and legislative changes governing the availability of agrichemicals for pest control. Novel crop production methods, which include genetic modification, must be considered for addressing these global food security issues. This project focuses on using one such method, CRISPR/Cas9, to alter the genome of the potato crop, Solanum tuberosum. The importance of potato cannot be overstated, being the third-largest food crop globally, and historically tied to Ireland. The rapidly changing climate and demand for increased yields calls for improvement to the potato, which historically has proven difficult to enhance, due to its tetraploid nature. The project specifically targeted the SGT3 gene, (Rhamnose:beta-solanine/beta-chaconine rhamnosyltransferase), a key gene involved in the production of solanine. The solanine production pathway is crucial to glycoalkaloid synthesis. While low levels of glycoalkaloids contribute to the flavour of potato, higher levels are toxic to humans and it is estimated that 14-27% of the U.S. potato crop is rejected due to dangerous glycoalkaloid levels. By inducing a small, 20 base-pair, deletion in the SGT3 gene, it is anticipated that its function would be disrupted, thus causing glycoalkaloid production to be supressed at an early stage of the pathway. In order to conduct this transformation in a “DNA-free” manner, a modified version of the CRISPR-Cas9 system was used, in which synthetic Cas9 endonuclease and synthetic sgRNA molecules were combined to form a ribonucleoprotien (RNP) complex. This RNP was then delivered to the plant tissue via particle bombardment. Following this, some plants were used for DNA analysis while others were subjected to regeneration protocols. A deletion in the SGT3 region was not detected in vivo, but was successfully digested in vitro, and important steps have been taken to ensuring this method will be more successful in the future. In addition, studies were conducted to examine how two cultivars of potato, Golden Wonder and Maris Piper, grow when exposed to different wavelengths of light and longer photoperiods. The strongest growth response was observed when plants were grown under various combinations of blue and red wavelengths. en
dc.format.mimetype application/pdf en
dc.language.iso en en
dc.publisher University College Cork en
dc.rights © 2020, Dara Eoin Meehan. en
dc.rights.uri https://creativecommons.org/licenses/by-nc-nd/4.0/ en
dc.subject CRISPR en
dc.subject Genetics en
dc.subject Plant biology en
dc.subject Plant genetics en
dc.subject Biology en
dc.title DNA-free genome editing of Solanum tuberosum. A CRISPR/Cas9-mediated proteolistic approach for novel crop production en
dc.type Masters thesis (Research) en
dc.type.qualificationlevel Masters en
dc.type.qualificationname MRes - Master of Research en
dc.internal.availability Full text not available en
dc.description.version Accepted Version en
dc.description.status Not peer reviewed en
dc.internal.school Biological, Earth and Environmental Sciences en
dc.internal.conferring Summer 2020 en
dc.availability.bitstream embargoed
dc.check.date 2023-05-13

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© 2020, Dara Eoin Meehan. Except where otherwise noted, this item's license is described as © 2020, Dara Eoin Meehan.
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