Mitochondrial membrane potential-based genome-wide RNAi screen of Trypanosoma brucei.
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.
Verner
Zdenek
Z
Biology Centre, Institute of Parasitology, Czech Academy of Sciences, and Faculty of Sciences, University of South Bohemia, 37005, Ceské Budejovice (Budweis), Czech Republic.
Paris
Zdenek
Z
Lukes
Julius
J
D000818:Animals; D005434:Flow Cytometry; D055785:Gene Knockdown Techniques / Q000379:methods*; D018503:Genome, Protozoan; D006801:Humans; D053078:Membrane Potential, Mitochondrial; D034741:RNA, Small Interfering / Q000235:genetics*; D014346:Trypanosoma brucei brucei / Q000235:genetics / Q000502:physiology*
D034741:RNA, Small Interfering