Latent transcriptional variations of individual Plasmodium falciparum uncovered by single-cell RNA-seq and fluorescence imaging

• DOI: 10.1371/journal.pgen.1008506
• Publication Date: 2019-12-19T18:47:23Z
• Authors: Katelyn A. Walzer; Hélène Fradin; Liane Y. Emerson; David L. Corcoran; Jen-Tsan Chi
• Keywords: 0301 basic medicine; Life Cycle Stages; 572; Sequence Analysis, RNA; Gene Expression Profiling; Plasmodium falciparum; Protozoan Proteins; Gene Expression Regulation, Developmental; QH426-470; Microfluidic Analytical Techniques; 3. Good health; 03 medical and health sciences; Multigene Family; Genetics; Single-Cell Analysis; In Situ Hybridization, Fluorescence; Research Article

Malaria parasites follow a complex life cycle that consists of multiple stages span from the human host to mosquito vector. Among species causing malaria, Plasmodium falciparum is most lethal, with clinical symptoms manifesting during intraerythrocytic developmental (IDC). During IDC, P. progresses through synchronous and continuous cascade transcriptional programming previously established using population analyses. While individual are known exhibit variations evade immune system or commit sexual fate, such rare expression heterogeneity largely undetectable on level. Therefore, we combined single-cell RNA-sequencing (scRNA-seq) microfluidic platform fluorescence imaging delineate among late asexual stages. The comparison between uncovered set undefined sex-specific genes. Asexual were segregated into three distinct clusters based differential genes encoding SERAs, rhoptry proteins, EXP2 plus transporters. Multiple pseudotime analyses revealed these stage-specific transitions distinct. RNA fluorescent in situ hybridization cluster-specific validated IDC defined highly variable pattern EXP2. Additionally, indicated huge transcript levels parasites. Overall, scRNA-seq RNA-FISH stage unexpected degrees potential impact regulation adaptive responses host.