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Různé metodické přístupy za účelem snížení exprese podjednotek komplexu I u hmyzích forem \kur{Trypanosoma brucei}
 HEROUTOVÁ, Barbora
Complex I (NADH:ubiquinone dehydrogenase) is the largest protein complex of the mitochondrial electron transport chain, but its presence and activity are not essential for the growth of two life cycle stages of Trypanosoma brucei. Here, we implemented various genetic methods to generate T. brucei cell lines that will lack or will have decreased levels of this complex. The methodological approaches included i) RNA interference of NDUFA6, a subunit predicted to be essential for the structural integrity, and of NUBM, a subunit essential for the complex I activity; ii) generation of double knock-out of NDUFA6 using CRISPR/Cas9; iii) generation of double knock-out of NUBM using homology replacement. These generated tools will help us to elucidate if this highly conserved complex is essential for other life cycle stages of this medically important parasite
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