Alternative approaches for control of gene expression in trypanosomatids

NIEMIROWICZ, G.T.; ALVAREZ, V.E.; BOUVIER, L.A.

Congreso; Molecular Parasitology Meeting XXXI; 2020

Regulatable gene expression in trypanosomatids relies on pre-establishing cell lines modified to express exogenous regulatory elements such asbacterial repressors and polymerases and therefore, inducible experimentation is frequently restricted to few laboratory strains. Moreover, extending thesemethodologies to other strains usually requires several additional transfection steps as well as lengthy periods in culture thus affecting the natural wild typeproperties sought after in field isolates.Here we explored alternative strategies for inducible transgene expression in Trypanosoma cruzi and Trypanosoma brucei. We studied the drugdependent modulation of GFP mRNA stability by insertion of a cis-acting riboswitch. This element consists of a Schistosoma mansoni hammerhead ribozymefused to a tetracycline aptamer. Reporter GFP expression could be downregulated by tetracycline addition, in T. cruzi epimastigotes as well as in procyclic T.brucei trypomastigotes. Thus, our results would suggest that these riboswitches could be used as alternative tools to study essential genes in trypanosomatids.