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Mitochondrial membrane potential-based genome-wide RNAi screen of Trypanosoma brucei

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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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Acknowledgments

We thank Paul T. Englund (Johns Hopkins University, Baltimore) for kindly providing the RNAi library and Piya Changmai for help with some experiments. This work was supported by the Ministry of Education of the Czech Republic (LC07032, 2B06129, and 6007665801) and the Praemium Academiae award to J.L.

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Authors and Affiliations

  1. Biology Centre, Institute of Parasitology, Czech Academy of Sciences, and Faculty of Sciences, University of South Bohemia, 37005, České Budějovice (Budweis), Czech Republic

    Zdeněk Verner, Zdeněk Paris & Julius Lukeš

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  1. Zdeněk Verner
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  2. Zdeněk Paris
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  3. Julius Lukeš
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Correspondence to Julius Lukeš.

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Zdeněk Verner and Zdeněk Paris contributed equally to this work.

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Verner, Z., Paris, Z. & Lukeš, J. Mitochondrial membrane potential-based genome-wide RNAi screen of Trypanosoma brucei . Parasitol Res 106, 1241–1244 (2010). https://doi.org/10.1007/s00436-010-1754-9

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  • DOI: https://doi.org/10.1007/s00436-010-1754-9

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