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.
authors: Whitehead Institute for Biomedical Research, Cambridge, Massachusetts, United States of America.
authors: Saima M
authors: SM
authors: Sidik
authors: Whitehead Institute for Biomedical Research, Cambridge, Massachusetts, United States of America.
authors: Caroline G
authors: CG
authors: Hackett
authors: School of Chemistry and Biomedical Sciences Research Complex, University of St. Andrews and EaStCHEM, North Haugh, St Andrews, Fife, Scotland, United Kingdom.
authors: Fanny
authors: F
authors: Tran
authors: School of Chemistry and Biomedical Sciences Research Complex, University of St. Andrews and EaStCHEM, North Haugh, St Andrews, Fife, Scotland, United Kingdom.
authors: Nicholas J
authors: NJ
authors: Westwood
authors: Whitehead Institute for Biomedical Research, Cambridge, Massachusetts, United States of America.
authors: Sebastian
authors: S
authors: Lourido
mesh_terms: D000953:Antigens, Protozoan / Q000737:chemistry / Q000235:genetics; D001483:Base Sequence; D064130:CRISPR-Associated Proteins; D064113:CRISPR-Cas Systems; D002460:Cell Line; D064112:Clustered Regularly Interspaced Short Palindromic Repeats; D059766:DNA End-Joining Repair; D023061:Gene Order; D018390:Gene Targeting / Q000379:methods; D005818:Genetic Engineering; D056426:Genetic Loci; D018503:Genome, Protozoan; D006801:Humans; D008969:Molecular Sequence Data; D010957:Plasmids / Q000235:genetics; D015800:Protozoan Proteins / Q000737:chemistry / Q000235:genetics; D016415:Sequence Alignment; D014122:Toxoplasma / Q000235:genetics*
chemical_list: D000953:Antigens, Protozoan; D064130:CRISPR-Associated Proteins; D015800:Protozoan Proteins; C047168:SAG1 antigen, Toxoplasma