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Public Library of Science, PLoS ONE, 6(9), p. e100450, 2014

DOI: 10.1371/journal.pone.0100450

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Efficient Genome Engineering of Toxoplasma gondii Using CRISPR/Cas9

Journal article published in 2014 by Saima M. Sidik, Caroline G. Hackett, Fanny Tran, Nicholas J. Westwood, Sebastian Lourido ORCID
This paper is made freely available by the publisher.
This paper is made freely available by the publisher.

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Abstract

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.