Characterization of RNA N6 adenosine methylation (m6A) in Plasmodium falciparum

Plasmodium falciparum, the causative agent of malaria, is a major health threat for nearly half of the world’s population. All symptoms of the disease arise during the 48 hour intra-erythtrocytic developmental cycle (IDC), during which the parasite replicates up to 32 daughter cells within human red blood cells. Underlying this reproductive cycle is a strictly timed pattern of gene expression which is regulated both on a transcriptional and post-transcriptional level. In this study, we characterized chemical modifications on messenger RNA as a novel layer of putative post-transcriptional gene regulation. We identified N6 adenosine methylation (m6A) to be the most abundant and highly dynamic mRNA modification, reflecting the parasites extreme transcriptomic adenosine content (~45%). We knocked down the putative m6A methyltransferase by CRISPR interference and combined directional RNA sequencing and m6A sequencing at three time points during the IDC to analyze the role of m6A on the fate of mRNA transcripts.