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Development of a CRISPR systemfor gene knockouts in Diplomonad parasites
Uppsala University, Disciplinary Domain of Science and Technology, Biology, Biology Education Centre.
2016 (English)Independent thesis Advanced level (professional degree), 20 credits / 30 HE creditsStudent thesis
Abstract [en]

This thesis describes the attempt to implement the CRISPR-Cas9 system in the two

diplomonad pathogens: Giardia intestinalis and Spironucleus salmonicida. Two approaches

of the implementation were done: first the Cas9 was expressed, followed by expression of the

guide RNA. Additionally, to extend the understanding of how small nuclear RNAs are

processed in Giardia intestinalis, two putative RNA processing enzymes were expressed and

localized within the cells. The results show expression of Cas9 in both G. intestinalis and S

.salmonicida. However, the Cas9 does not translocate to the nuclei, which is crucial for a

functional gene knockout system. The approach where the guide RNA is expressed show a

successful expression. In the experiments with the two putative RNA processing enzymes

expression is observed in both cases, where one of the enzymes is localized to the nuclei.

Place, publisher, year, edition, pages
UPTEC X, 16 006
National Category
Microbiology Engineering and Technology
URN: urn:nbn:se:uu:diva-294365OAI: diva2:929545
Educational program
Molecular Biotechnology Engineering Programme
Available from: 2016-05-19 Created: 2016-05-19 Last updated: 2016-05-19Bibliographically approved

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