CRISPR/Cas9 mediated deletion of genes encoding putative cell cycle regulators in Plasmodium falciparum

• Main Author: Von Grüning, Hilde
• Other Authors: Birkholtz, Lyn-Marie
• Language: en
• Published: University of Pretoria 2018
• Subjects: UCTD
• Online Access: http://hdl.handle.net/2263/65954; Von Grüning, H 2017, CRISPR/Cas9 mediated deletion of genes encoding putative cell cycle regulators in Plasmodium falciparum, MSc Dissertation, University of Pretoria, Pretoria, viewed yymmdd <http://hdl.handle.net/2263/65954>

Functional genomic tools can be used to interrogate unique features of parasite biology that could be marked for future intervention strategies. One such unique biological feature is the atypical cell cycle of the Plasmodium falciparum parasite, with a particular interest in the key regulators that modulate the parasite’s cell cycle progression. The orderly progression of the cell cycle of the parasite allows it to undergo a vast expansion of parasite numbers during a malaria infection and cause pathologies in infected individuals. However, the precise control mechanisms and functional cascades involved in the P. falciparum parasite’s unusual cell cycle have not yet been fully elucidated. A recent study showed that gene expression profiles of the parasite switch between states of cell cycle arrest (quiescence) to cell cycle re-entry (proliferation). Global transcriptome expression data from this study was used to identify potential cell cycle regulators. The study identified a number of putative cell cycle regulators and their predicted functional interacting partners, particularly transcription factors and several members of the origin of replication complex. The study subsequently aimed to functionally validate these putative regulators in a reverse genetics approach by creating targeted disruption of the putative regulatory genes using the CRISPR-Cas9 system. Preliminary knockout studies indicated possible essential phenotypes of these putative cell cycle regulators in P. falciparum parasites. Given that cell cycle elements of the P. falciparum parasites are unique and divergent from those of the human host, the information provided in this study dissects the role of regulators in cell cycle modulation in the parasite. Not only is this important to understand parasite biology, but these cell cycle regulators are attractive potential sites for chemical interference of parasite proliferation and thereby provide as novel drug targets for antimalarial discoveries. === Dissertation (MSc)--University of Pretoria, 2017. === Biochemistry === MSc === Unrestricted