Efficient genome engineering of Toxoplasma gondii using CRISPR/Cas9
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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.
Sidik , S M , Hackett , C G , Tran , F , Westwood , N J & Lourido , S 2014 , ' Efficient genome engineering of Toxoplasma gondii using CRISPR/Cas9 ' PLoS One , vol 9 , no. 6 , e100450 . DOI: 10.1371/journal.pone.0100450
© 2014 Sidik et al. This is an open-access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
This work was supported in part by NIH grant 1DP5OD017892 to S.L
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