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Mitochondrial membrane potential-based genome-wide RNAi screen of Trypanosoma brucei
- Verner, Zdeněk, Paris, Zdeněk, Lukeš, Julius
- Parasitology research 2010 v.106 no.5 pp. 1241-1244
- RNA interference, Trypanosoma brucei, clones, dyes, flow cytometry, membrane potential, screening
- We have screened the Trypanosoma brucei genome-wide RNAi library by staining the procyclics with the dye JC-1 followed by sorting the differentially stained cells by flow cytometry. This allowed us to highly enrich for cells in which mitochondrial membrane potential was decreased. We have further validated a subset of selected clones by a reverse approach in which we showed that cloning the selected genomic regions into another RNAi vector also results in a drop in mitochondrial membrane potential.