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In  Trypanosoma brucei , Argonaute1 ( Tb AGO1) is an essential component of the RNA interference (RNAi) pathway. While characterizing a  TbAGO1  conditional knockout cell line, we discovered that, upon blockage of  TbAGO1  transcription, the RNAi response to transfected double-stranded RNA was severely inhibited, although there was no change in the  Tb AGO1 protein level. This observation suggested that steady-state  Tb AGO1 was not sufficient to fully support the RNAi response to transfected dsRNA and implicated newly synthesized Argonaute in this phenomenon. Indeed, a translational blockade of  TbAGO1  mRNA with an antisense morpholino oligonucleotide resulted in inhibition of the RNAi response, even though the steady-state level of  Tb AGO1 remained unchanged during the time of the assay. Thus, we concluded that in  T. brucei , newly synthesized  Tb AGO1 is required to support an efficient RNAi response. We speculate that newly processed siRNAs may be preferentially loaded onto newly synthesized  Tb AGO1, and this mechanism may contribute to the homeostasis of the RNAi pathway.

Depletion of newly synthesized Argonaute1 impairs the RNAi response in Trypanosoma brucei