Phenotypic activation and repression of three ELAV-like proteins in Trypanosoma brucei. Trypanosoma brucei

Three potential ELAV-like proteins of T. brucei, including Tb927.3.2930, Tb927.7.5380, and Tb927.8.6650, were either inhibited by RNAi or phenotypically activated by over-expression, followed by microarray analysis of the transcriptome. The results indicated that these ELAV-like proteins regulate the abundance of a large number of T. brucei transcripts, potentially through regulation of mRNA stability. Overall design: Each of the three ELAV-like proteins were either inhibited by RNAi or over-expressed, in stable transgenic procyclic form cell lines. Total RNA was extracted 48h after tetracycline induction of the constructs (except for total RNA from Tb927.8.6650 RNAi which was extracted 24h after tet-induction). cDNA was synthesized using TaqMan Reverse Transcription Reagents with random hexamer priming (Applied Biosystems), and sent to NimbleGen for labeling and hybridization. Non-induced cells were analyzed in parallel.