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Targeting prolyl-tRNA synthetase via a series of ATP-mimetics to accelerate drug discovery against toxoplasmosis

ABSTRACT: To shed light on the parasiticidal mechanisms of L35, we have adapted a workflow which combines a forward genetic approach based on transcriptome sequencing, computational mutation discovery, and CRISPR/Cas9 genome editing in Toxoplasma gondii. Drug-resistant parasites were generated by chemical random mutagenesis. Multiple independent resistant lines were isolated. Single nucleotide variations (SNVs) were identified based on NGS transcriptomic analysis. By focusing on mutations present in coding sequences, we identified a single gene, TgPRS, that harbored SNVs leading to amino acid substitutions in the 6 drug-resistant lines obtained that were not present in the parental strain. Finally, using CRISPR/Cas9 genome editing we confirmed that the mutations identified confer resistance against L35.

ORGANISM(S): Toxoplasma gondii RH Homo sapiens

PROVIDER: GSE217090 | GEO | 2023/01/22

REPOSITORIES: GEO

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