Efficient genome engineering of Toxoplasma gondii using CRISPR/Cas9

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

doi:10.1371/journal.pone.0100450

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 journal › Article › peer-review

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.

Original language	English
Article number	e100450
Number of pages	8
Journal	PLoS One
Volume	9
Issue number	6
DOIs	https://doi.org/10.1371/journal.pone.0100450
Publication status	Published - 27 Jun 2014

Keywords

Invasion
Systems
Marker
Genes

UN SDGs

This output contributes to the following UN Sustainable Development Goals (SDGs)

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10.1371/journal.pone.0100450

Westwood_2014_PLOSone_Efficient
© 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.
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title = "Efficient genome engineering of Toxoplasma gondii using CRISPR/Cas9",

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.",

keywords = "Invasion, Systems, Marker, Genes",

author = "Sidik, {Saima M.} and Hackett, {Caroline G.} and Fanny Tran and Westwood, {Nicholas J.} and Sebastian Lourido",

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AU - Sidik, Saima M.

AU - Hackett, Caroline G.

AU - Tran, Fanny

AU - Westwood, Nicholas J.

AU - Lourido, Sebastian

N1 - This work was supported in part by NIH grant 1DP5OD017892 to S.L

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N2 - 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.

AB - 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.

KW - Invasion

KW - Systems

KW - Marker

KW - Genes

U2 - 10.1371/journal.pone.0100450

DO - 10.1371/journal.pone.0100450

M3 - Article

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JO - PLoS One

JF - PLoS One

IS - 6

M1 - e100450

ER -

Efficient genome engineering of Toxoplasma gondii using CRISPR/Cas9

Abstract

Keywords

UN SDGs

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