Toxoplasma gondii is an intracellular parasite of the Apicomplexa phylum. It is the causative agent of toxoplasmosis. T. gondii, along with other Apicomplexa, possesses a relict plastid named the apicoplast. This plastid houses several essential metabolic pathways. The importance of the apicoplast and its evolutionary divergence from the host make it an attractive drug target. Despite a thorough understanding of its metabolic pathways, the transporter proteins involved in metabolite shuttling from and to the apicoplast are largely unknown. This master thesis focused on the identification and characterization of 5 new apicoplast transporters, 3 belonging to the MFS superfamily (MFS5, MFS11, and MFS12) and 2 permeases (APE1 and APE2). Using inducible knockdown systems, the 3 MFS proved to be essential for parasite survival while the 2 permeases were found to be dispensable. Among the MFS, only MFS11 was found to be dispensable for apicoplast housekeeping, with parasites keeping the organelle after its downregulation. Instead, MFS5 and MFS12 are likely involved in apicoplast housekeeping and cause loss of the organelle, resulting in the delayed death phenomenon, upon their downregulation. Although the transporters’ substrates have not been definitively determined, untargeted metabolomics revealed GMP and AMP as candidate substrates for MFS11.