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Published on: July 22, 2014
Mitochondrial membrane potential-based genome-wide RNAi screen of Trypanosoma brucei
Zdenek Verner1, Zdenek Paris, Julius Lukes
1Biology Centre, Institute of Parasitology, Czech Academy of Sciences, and Faculty of Sciences, University of South Bohemia, 37005, Ceské Budejovice (Budweis), Czech Republic.
Abstract:
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.
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