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Functional Characterization of Dense Granule Proteins in
Meng-Jie Bai1, Jin-Lei Wang1, Hany M Elsheikha2
1State Key Laboratory of Veterinary Etiological Biology, Key Laboratory of Veterinary Parasitology of Gansu Province, Lanzhou Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Lanzhou, China.
Frontiers in Cellular and Infection Microbiology
|September 14, 2018
Summary
Researchers disrupted 17 dense granule protein (GRA) genes in Toxoplasma gondii. Deleting these GRA genes did not impact parasite growth, host cell egress, or virulence in mice, suggesting other roles in different parasite strains or stages.
Area of Science:
- Parasitology
- Molecular Biology
- Infectious Diseases
Background:
- Toxoplasma gondii infection is a significant public health concern.
- Dense granule proteins (GRAs) are crucial for T. gondii invasion and replication.
- The precise functions of several GRAs are not fully understood.
Purpose of the Study:
- To investigate the roles of 17 dense granule proteins (GRAs) in T. gondii pathogenesis.
- To determine the necessity of these GRAs for parasite growth, host cell egress, and virulence.
Main Methods:
- CRISPR-Cas9 gene editing was used to create single knockout mutants for 17 GRA genes in the T. gondii RH strain.
- In vitro assays included plaque assays and egress assays to assess parasite growth and egress.
- In vivo studies involved infection of BALB/c mice to evaluate parasite virulence.
Main Results:
- CRISPR-Cas9 successfully abolished the expression of the targeted 17 GRA genes.
- Deletion of these 17 GRA genes did not significantly affect in vitro parasite growth or egress from host cells.
- The mutant strains exhibited no significant reduction in virulence in a mouse model of infection.
Conclusions:
- The investigated 17 GRAs are not essential for the in vitro growth or mouse virulence of the T. gondii RH strain.
- These GRAs may possess distinct functions in different T. gondii genotypes or life cycle stages.
- Further research is needed to elucidate the roles of these GRAs in other contexts.
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