Efficient genome engineering of Toxoplasma gondii using CRISPR/Cas9

Saima M. Sidik, Caroline G. Hackett, Fanny Tran, Nicholas J. Westwood, Sebastian Lourido*

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review

185 Citations (Scopus)
3 Downloads (Pure)


Toxoplasma gondii is a parasite of humans and animals, and a model for other apicomplexans including Plasmodium spp., the causative agents of malaria. Despite many advances, manipulating the T. gondii genome remains labor intensive, and is often restricted to lab-adapted strains or lines carrying mutations that enable selection. Here, we use the RNA-guided Cas9 nuclease to efficiently generate knockouts without selection, and to introduce point mutations and epitope tags into the T. gondii genome. These methods will streamline the functional analysis of parasite genes and enable high-throughput engineering of their genomes.

Original languageEnglish
Article numbere100450
Number of pages8
JournalPLoS One
Issue number6
Publication statusPublished - 27 Jun 2014


  • Invasion
  • Systems
  • Marker
  • Genes


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