Whole genome sequencing of Leishmania donovani AAP3 mutants

After infecting host macrophages, Leishmania parasites encounter arginine deprivation. This activates an arginine deprivation response (ADR) pathway, resulting in upregulating the abundance and activity of the parasite arginine transporter (AAP3). We used whole genome sequencing (WGS) to determine the genotype of several mutants following CRISPR/Cas9-mediated disruption of the AAP3 locus. Mutants that retained only a single AAP3 allele showed a basal level of arginine transport but lacked the ability to respond to arginine starvation. While these mutants grow normally in culture, they were impaired in their ability to develop inside THP-1 macrophages and less infective in BALB/c mice.