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RNAi libraries for high throughput phenotyping and drug resistance screening in the African trypanosome

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African trypanosomes are major pathogens of humans and livestock and represent a model for studies of unusual protozoal biology. We developed a high throughput phenotyping approach termed RNA interference (RNAi) target sequencing, or RIT -seq that, using Illumina sequencing, maps fitness-costs associated with RNAi. Analysis of bloodstream and insect life-cycle stages and differentiated libraries linked thousands of previously uncharacterised and ‘hypothetical’ genes to essential functions. The data-set also provides much needed genetic validation to identify new drug-targets. In a second application, all five current anti-trypanosomal drugs were used for RIT -seq screens, revealing the transporters, organelles, enzymes and metabolic pathways that function to facilitate anti-trypanosomal drug action.

This talk is part of the Seminars on Quantitative Biology @ CRUK Cambridge Institute series.

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