Talented post-doctoral researchers and PhD candidates, who are enthusiastic about using experimental genetics approaches to study the molecular and cellular biology of the malaria parasite, are encouraged to contact Taco to explore fellowship opportunities.
Motivated MSc students with an interest in an unusual but medically extremely relevant eukaryotic microbe, who would like to learn generally applicable molecular biology and microscopy techniques, should also contact Taco. There are a number of suitable projects, usually for a period of ≥6 months (see below for a general project outline).
Masters project
Studying malaria parasite biology during host switching using novel genetic tools
Malaria is a devastating disease of unquestioned human suffering in largely poorer countries. However, it is far less well known that the malaria parasite is also a captivating, unique, and extremely useful model organism that allows us to study general principles of molecular and cell biology as well as unique parasite-specific features. In our lab, we develop and employ experimental genetics tools combined with advanced flow cytometric, proteomic, and microscopical techniques to shed light on fundamental parasite biology in particular during host switching, strengthening the quest for prophylactic and transmission-blocking intervention strategies. One of our key interests is the functioning of the mitochondrion. Malaria parasites only have a single, abnormal mitochondrion, target of the drug atovaquone [1]. Interestingly, the mitochondrion changes dramatically in morphology and function during transmission [2,3,4].
All current projects will make use of in vitro cultured human malaria parasites (Plasmodium falciparum). Using novel experimental genetic approaches including CRISPR-Cas-based methods, it is possible to study functioning of the malaria parasite mitochondrion or specific proteins or protein complexes. This could include the generation and study of loss-of-function mutants, conditional knockdowns, and spatio-temporal analysis of fluorescently labelled proteins. Current topics include mitochondrial transport proteins [5], proteins involved in mitochondrial fission [6], and various mitochondrial protein complexes involved in oxidative phosphorylation [7] or cristae biogenesis.
Most projects will begin with a bioinformatic evaluation of the chosen target(s), followed by the design and generation of the required transfection vectors using standard molecular biology techniques. Next, you will learn to grow, transfect, and monitor the parasites. The exact methods used in the subsequent functional characterization of the generated mutant parasites are largely dependent on the specific project but will usually include specialised culturing, light and fluorescent microscopy, flow cytometry, and other molecular methods.
Objectives of this Masters project are to generate genetically modified parasite lines to study the molecular mechanisms of host switching and analyse life-cycle progression and cell biology of the generated mutants with a focus on transmission stages.
Techniques to be learned and used in this project include: database mining and in silico design of transfection vectors; standard molecular biological techniques (including PCR, plasmid cloning, gDNA isolation); Western blot analysis; brightfield and fluorescence microscopy (of various malaria parasite life cycle stages); in vivo or in vitro cultures and assays to monitor and characterize asexual- and sexual-stage development. And possibly: flow cytometric analysis of parasite growth; super-resolution microscopy, nitrogen cavitation, biochemical assays.
PMIDs: [1] 19575561, [2] 25831536, [3] 25843709, [4] 29669282, [5] 26796412, [6] 34425697, [7] 34155201.
Taco Kooij 