Efficient genome engineering of Toxoplasma gondii using CRISPR/Cas9

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 mu...

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Bibliographic Details
Published in:PloS one 2014-06, Vol.9 (6), p.e100450-e100450
Main Authors: Sidik, Saima M, Hackett, Caroline G, Tran, Fanny, Westwood, Nicholas J, Lourido, Sebastian
Format: Article
Language:English
Subjects:
Antigens, Protozoan - chemistry
Antigens, Protozoan - genetics
Base Sequence
Biology and Life Sciences
Biomedical research
Cell Line
Cloning
Clustered Regularly Interspaced Short Palindromic Repeats
CRISPR
CRISPR-Associated Proteins
CRISPR-Cas Systems
Deoxyribonucleic acid
Dihydrofolate reductase
DNA
DNA End-Joining Repair
Engineering
Epitopes
Functional analysis
Gene loci
Gene Order
Gene Targeting - methods
Genetic Engineering
Genetic Loci
Genome editing
Genome, Protozoan
Genomes
Humans
Kinases
Malaria
Molecular Sequence Data
Mutation
Nuclease
Parasites
Plasmids - genetics
Proteins
Protozoan Proteins - chemistry
Protozoan Proteins - genetics
Ribonucleic acid
RNA
Science
Sequence Alignment
Toxoplasma - genetics
Toxoplasma gondii
Vector-borne diseases
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Description
Summary: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.
ISSN:1932-6203
1932-6203
DOI:10.1371/journal.pone.0100450