RNA sequencing of Rbpm1 knockout in Plasmodium yoelii. RNA sequencing of Rbpm1 knockout in Plasmodium yoelii

RBPm1, an RNA-binding protein, plays a crucial role in regulating axoneme assembly during male gametogenesis of Plasmodium yoelii. To elucidate the molecular mechanisms that underlie this process, we conducted RNA-seq analysis to investigate changes in the male transcriptome resulting from the loss of RBPm1. Overall design: To obtain highly pure male gametocytes, we employed a DFsc7 reporter strain that selectively exhibited GFP expression within these cells. We created a mutant strain by deleting RBPm1 from the DFsc7 strain. Male gametocytes from both the DFsc7 and mutant strains were sorted using fluorescence-activated cell sorting (FACS), relying on GFP fluorescence intensity as a criterion, and subsequently subjected to RNA-seq analysis. Addition: Through Principal component analysis (PCA), it was observed that the RNA-seq data from the initial sample, WT biol rep1, exhibited poor reproducibility. Subsequently, we re-sorted and obtained three additional WT samples (WT_biol_rep4, WT_biol_rep5, and WT_biol_rep6) for RNA-seq, resulting in improved data reproducibility.