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    CRISPR/Cas9-mediated Viral Interference in Plants

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    Manal Tashkandi Dissertation.pdf
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    Description:
    Manal Tashkandi Dissertation
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    Type
    Dissertation
    Authors
    Tashkandi, Manal cc
    Advisors
    Mahfouz, Magdy M. cc
    Committee members
    Blilou, Ikram cc
    Ghaffour, NorEddine cc
    Benhamed, Moussa
    Program
    Bioscience
    KAUST Department
    Biological and Environmental Sciences and Engineering (BESE) Division
    Date
    2018-05
    Permanent link to this record
    http://hdl.handle.net/10754/628029
    
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    Abstract
    In prokaryotes, CRISPR/Cas9 system provides molecular immunity to bacteria and archaea against invading phages, conjugative plasmids and nucleic acids. CRISPR/Cas9 system has been adapted for targeted genome editing across diverse eukaryotic species for a variety of applications in basic and applied research. In this dissertation, I propose to adapt the CRISPR/Cas9 system to function as molecular immunity machinery against plant DNA viruses. Therefore, to test whether the CRISPR/Cas9 system is portable to plants, I produced plants stably over-expressing Cas9 and sgRNAs against single or multiple DNA viruses in Nicotiana benthamiana (N. benthamiana) and tomato (Solanum lycopersicum) plants. sgRNAs targeting the Cas9 endonuclease against different coding and non-coding viral sequences were tested in virus- interference experiments. I explored the possibility of generating robust interference against single and multiple DNA viruses. Subsequently, I studied the possibility of virus evasion of the CRISPR/Cas9 machinery and evolution of the virus escapees. Finally, I produced N. benthamaiana and tomato plants stably expressing the CRISPR/Cas9 machinery for developing durable virus resistance. Furthermore, developing effective viral-interference system in plants will help to understand the molecular underpinning of virus biology and host-defense mechanisms against plant viruses. In conclusion, my research project attempted to establish the efficacy and extend the utility of CRISPR/Cas9 system for viral interference in plants which promise exciting applications including producing engineered plants resistant to multiple viral infection.
    DOI
    10.25781/KAUST-RDG1N
    ae974a485f413a2113503eed53cd6c53
    10.25781/KAUST-RDG1N
    Scopus Count
    Collections
    Biological and Environmental Sciences and Engineering (BESE) Division; Bioscience Program; Dissertations

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