One at a time: dissecting the spatial interactome in African Trypanosomes

FUNDING: 9 weeks (full time, 37 hrs per week, £10.57 per hour, £690 consumables)
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Dr Lianne Lansink, Biology, York

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Antigenic variation, the expression of a single antigen out of a large gene family, combined with periodic antigen switching, is a sophisticated virulence mechanism for immune evasion in African trypanosome parasites. These parasites express a single variant-surface-glycoprotein (VSG) from a vast genetic repertoire (>2600 genes). Recent research has uncovered a range of subnuclear bodies specifically organised to transcribe and process VSG transcripts. Thus far, only three proteins have been found that specifically inhabit these bodies and play a role in VSG gene regulation. To identify other regulators, we have performed proximity labelling (TurboID) combined with proteomics and currently have a list of candidate genes. In this project the candidate genes will be endogenously tagged, and the resulting fusion proteins analysed by western blot and fluorescence microscopy. From those, proteins that uniquely localise to the nuclear bodies of interest will be prioritised for depletion (RNAi & CRISPR/Cas9) studies to study their role on VSG expression control (RNA-Seq). The successful candidate will culture and genetically modify Trypanosoma brucei parasites (class-2 pathogens), learn general molecular biology techniques, and immunofluorescence. Students will be expected to present their findings orally at a research day in York in September 2023.