AdvisorsMahfouz, Magdy M.
KAUST DepartmentBiological and Environmental Science and Engineering (BESE) Division
Embargo End Date2019-06-27
Permanent link to this recordhttp://hdl.handle.net/10754/628029
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Access RestrictionsAt the time of archiving, the student author of this dissertation opted to temporarily restrict access to it. The full text of this dissertation became available to the public after the expiration of the embargo on 2019-06-27.
AbstractIn 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.
CitationTashkandi, M. (2018). CRISPR/Cas9-mediated Viral Interference in Plants. KAUST Research Repository. https://doi.org/10.25781/KAUST-RDG1N