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PLoS One. 2014 Jun 27;9(6):e100450. doi: 10.1371/journal.pone.0100450. eCollection 2014.

Efficient genome engineering of Toxoplasma gondii using CRISPR/Cas9.

Author information

1
Whitehead Institute for Biomedical Research, Cambridge, Massachusetts, United States of America.
2
School of Chemistry and Biomedical Sciences Research Complex, University of St. Andrews and EaStCHEM, North Haugh, St Andrews, Fife, Scotland, United Kingdom.

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.

PMID:
24971596
PMCID:
PMC4074098
DOI:
10.1371/journal.pone.0100450
[Indexed for MEDLINE]
Free PMC Article

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