Trypanosoma brucei RNA modifications
The goal of the project is to map the positions of 5-methylcytosine in Trypanosoma brucei RNA and identify the responsible enzymes necessary for 5-methylcytosine formation. T. brucei RNA was isolated, treated with sodium bisulfite, and analyzed by next generation sequencing. Cytosines will be read as Ts whereas 5-methylcytosines will be read as Cs after bisulfite treatment and sequencing. Sequencing was from total RNA and small RNA. In some experiments, sequencing was performed on RNA isolated from uninduced and induced RNA methyltransferase RNAi mutants.