A Plasmodium knowlesi A1-H.1 transcriptome time course focusing on the late asexual blood stages

The zoonotic parasite Plasmodium knowlesi is closely related to Plasmodium vivax , the second leading cause of human malaria. P. knowlesi line A1-H.1 can be maintained in human erythrocytes and is an experimental model for P. vivax biology. We present the first transcriptome time course of the intraerythrocytic developmental cycle (IDC) of P. knowlesi A1-H.1 grown in human normocytes. Bulk RNA-sequencing was performed at five IDC stages using tightly synchronised cultures, enabling identification of constitutively expressed genes and stage-specific biomarkers, and investigation of temporal expression patterns of multigene families. Comparative analysis with P. vivax orthologues revealed strong genome-wide conservation of temporal expression. Analysis of invasion-associated genes and ApiAP2 transcription factors identified genes with conserved expression patterns for which P. knowlesi is likely a good model to study the P. vivax gene. To support comparative analyses, we developed an open-access interactive webtool to explore and visualise P. knowlesi – P. vivax – P. falciparum orthologue expression across the IDC ( https://interactive.itg.be/app/pk-orthologue-expression-viewer ). This time course dataset provides a reference transcriptome framework for P. knowlesi A1-H.1 and a resource for comparative Plasmodium biology. The interactive webtool facilitates rapid identification of candidate genes with conserved temporal expression, streamlining functional studies in which P. knowlesi A1-H.1 serves as a model for P. vivax .