The kinetoplastid parasite Leishmania infantum is responsible for zoonotic visceral leishmaniasis in the mediterranean basin, where dogs are the reservoir. Differential gene expression analysis of metacyclic promastigotes in axenic culture by whole genome DNA microarray hybridization revealed up-regulation of two unidentified genes that are absent in the parasite's genome databases. Sequence analysis has revealed that these genes encode for guide RNAs (gRNAs), which are located in the kinetoplast and participate in the kinetoplastid-specific uridine insertion/deletion RNA editing process. Northern blot assays have confirmed that both gRNA genes are up-regulated in metacyclic promastigotes, thus suggesting that uridine insertion/deletion RNA editing contributes to metabolic shifts at this stage. A screening strategy described herein has revealed an uncharacterized 16S-like rRNA transcript as a target of one of the aforementioned gRNAs.
Keywords: Guide RNA; Kinetoplastids; Leishmania infantum; Metacyclic promastigotes; Minicircles; Peanut agglutinin; RNA-editing.
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