Differentiation between distinct stages is fundamental for the life cycle of intracellular protozoan parasites and for transmission between hosts, requiring stringent spatial and temporal regulation. Here, we apply kinome-wide gene deletion and gene tagging in Leishmania mexicana promastigotes to define protein kinases with life cycle transition roles. Whilst 162 are dispensable, 44 protein kinase genes are refractory to deletion in promastigotes and are likely core genes required for parasite replication. Phenotyping of pooled gene deletion mutants using bar-seq and projection pursuit clustering reveal functional phenotypic groups of protein kinases involved in differentiation from metacyclic promastigote to amastigote, growth and survival in macrophages and mice, colonisation of the sand fly and motility. This unbiased interrogation of protein kinase function in Leishmania allows targeted investigation of organelle-associated signalling pathways required for successful intracellular parasitism.

• MeSH
• Models, Biological MeSH
• Cell Differentiation * MeSH
• CRISPR-Cas Systems genetics   MeSH
• Gene Deletion MeSH
• Flagella enzymology   MeSH
• Leishmania mexicana cytology   enzymology   MeSH
• Leishmaniasis parasitology   pathology   MeSH
• Mutation genetics   MeSH
• Mice, Inbred BALB C MeSH
• CRISPR-Associated Protein 9 metabolism   MeSH
• Protein Kinases genetics   metabolism   MeSH
• Proteome metabolism   MeSH
• Psychodidae parasitology   MeSH
• Cell Survival MeSH
• Animals MeSH
• Check Tag
• Female MeSH
• Animals MeSH
• Publication type
• Journal Article MeSH
• Research Support, Non-U.S. Gov't MeSH