ArticleInfection and immunity2016
Analysis of Noncanonical Calcium-Dependent Protein Kinases in Toxoplasma gondii by Targeted Gene Deletion Using CRISPR/Cas9.
Article in Infection and immunity, 2016. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 43 papers.
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Who cites it
43 citing papers in PubMed.
- Calcium-dependent protein kinases 2A involved in the growth of both asexual and sexual stages of Cryptosporidium parvum.PLoS neglected tropical diseases · 2025Article
- Impact of sodium alginate hydrogel containing bacteriophage peptides that specifically bind to the EtCab protein on the inhibition of Eimeria tenella infection.Veterinary research · 2025Article
- Functional dissection of prenyltransferases reveals roles in endocytosis and secretory vacuolar sorting in type 2-ME49 strain ofVirulence · 2024Article
- PP1 phosphatase controls both daughter cell formation and amylopectin levels in Toxoplasma gondii.PLoS biology · 2024Article
- TBC9, an essential TBC-domain protein, regulates early vesicular transport and IMC formation in Toxoplasma gondii.Communications biology · 2024Article
- TheeLife · 2024Article
- The plant-like protein phosphatase PPKL regulates parasite replication and morphology in Toxoplasma gondii.Parasites & vectors · 2024Article
- Functional screening revealsmBio · 2023Article
- Article
- The Toxoplasma micropore mediates endocytosis for selective nutrient salvage from host cell compartments.Nature communications · 2023Article
- A positive feedback loop mediates crosstalk between calcium, cyclic nucleotide and lipid signalling in calcium-induced Toxoplasma gondii egress.PLoS pathogens · 2022Article
- Paving the Way: Contributions of Big Data to Apicomplexan and Kinetoplastid Research.Frontiers in cellular and infection microbiology · 2022Review
- Identification of new components of the basal pole ofFrontiers in cellular and infection microbiology · 2022Article
- Anti-Toxoplasma gondii effect of lupane-type triterpenes from the bark of black alder (Alnus glutinosa) and identification of a potential target by reverse docking.Parasite (Paris, France) · 2022Article
- Biotinylation of the Neospora caninum parasitophorous vacuole reveals novel dense granule proteins.Parasites & vectors · 2021Article
- Cryo-EM structure of cortical microtubules from human parasite Toxoplasma gondii identifies their microtubule inner proteins.Nature communications · 2021Article
- Protein kinases in Toxoplasma gondii.International journal for parasitology · 2021Review
- Functional Characterization of 17 Protein Serine/Threonine Phosphatases inFrontiers in cell and developmental biology · 2021Article
- Auranofin Resistance inFrontiers in cellular and infection microbiology · 2021Article
- Study on Circulating Antigens in Serum of Mice With Experimental Acute Toxoplasmosis.Frontiers in microbiology · 2020Article
Corrections and comments
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Authors and funding
3 authors.
Funding
Abstract
Calcium-dependent protein kinases (CDPKs) are expanded in apicomplexan parasites, especially in Toxoplasma gondii where 14 separate genes encoding these enzymes are found. Although previous studies have shown that several CDPKs play a role in controlling invasion, egress, and cell division in T. gondii, the roles of most of these genes are unexplored. Here we developed a more efficient method for gene disruption using CRISPR (clustered regularly interspaced short palindromic repeats)/Cas9 (CRISPR-associated protein 9) that was modified to completely delete large, multiexonic genes from the genome and to allow serial replacement by recycling of the selectable marker using Cre-loxP. Using this system, we generated a total of 24 mutants in type 1 and 2 genetic backgrounds to ascertain the functions of noncanonical CDPKs. Remarkably, although we were able to confirm the essentiality of CDPK1 and CDPK7, the majority of CDPKs had no discernible phenotype for growth in vitro or infection in the mouse model. The exception to this was CDPK6, loss of which leads to reduced plaquing, fitness defect in a competition assay, and reduced tissue cyst formation in chronically infected mice. Our findings highlight the utility of CRISPR/Cas9 for rapid serial gene deletion and also suggest that additional models are needed to reveal the functions of many genes in T. gondii.
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