Evaluation of the basic functions of six calcium-dependent protein kinases in Toxoplasma gondii using CRISPR-Cas9 system
| العنوان: | Evaluation of the basic functions of six calcium-dependent protein kinases in Toxoplasma gondii using CRISPR-Cas9 system |
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| المؤلفون: | Kai Chen, Xing-Quan Zhu, Jin-Lei Wang, Si-Yang Huang, Hong-Rui Ning, Ting-Ting Li |
| المصدر: | Parasitology Research. 115:697-702 |
| بيانات النشر: | Springer Science and Business Media LLC, 2015. |
| سنة النشر: | 2015 |
| مصطلحات موضوعية: | 0301 basic medicine, Genotype, CRISPR-Associated Proteins, Virulence, Gene Knockout Techniques, Mice, 03 medical and health sciences, Drug Delivery Systems, Plasmid, parasitic diseases, Animals, Humans, CRISPR, Clustered Regularly Interspaced Short Palindromic Repeats, Gene, Mice, Inbred BALB C, General Veterinary, biology, Kinase, Toxoplasma gondii, General Medicine, biology.organism_classification, Virology, Phenotype, Specific Pathogen-Free Organisms, 030104 developmental biology, Infectious Diseases, Lytic cycle, Insect Science, Calcium, Female, Parasitology, CRISPR-Cas Systems, Protein Kinases, Toxoplasma, Plasmids |
| الوصف: | Toxoplasma gondii, an important protozoan parasite, infects almost all warm-blooded animals and humans. Although treatments in T. gondii are limited by the lack of effective drugs, some calcium-dependent kinases were demonstrated as the promising drug targets to chemotherapy against T. gondii due to their essential roles in T. gondii and absence from their hosts. The objectives of the present study were to investigate the functions of six calcium-dependent protein kinases (CDPK4, CDPK4A, CDPK5, CDPK6, CDPK8, and CDPK9) in T. gondii to assess whether they are suitable for designing as drug targets. We used the CRISPR-Cas9 system to disrupt six CDPK genes successfully by insertion of DHFR* at the guide RNA-targeted region in the six endogenous CDPK loci and successfully obtained the six knockout (KO)-CDPK strains. The biological characteristics of the six strains were evaluated by plaque assays, invasion, egress, replication, and virulence assays, respectively. The results indicated that there was no significant difference between the six KO-CDPK strains and wild-type strain in virulence and the lytic cycle including invasion, egress, and replication. The conclusion was the six CDPKs are not essential for T. gondii lytic cycle and also not virulence factors for mice, suggesting that the six CDPKs may participate in other functions in T. gondii. |
| تدمد: | 1432-1955 0932-0113 |
| الوصول الحر: | https://explore.openaire.eu/search/publication?articleId=doi_dedup___::d01c11c6b697fa40827ebe1d789e5735Test https://doi.org/10.1007/s00436-015-4791-6Test |
| حقوق: | CLOSED |
| رقم الانضمام: | edsair.doi.dedup.....d01c11c6b697fa40827ebe1d789e5735 |
| قاعدة البيانات: | OpenAIRE |
| تدمد: | 14321955 09320113 |
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