Evaluation of the basic functions of six calcium-dependent protein kinases in Toxoplasma gondii using CRISPR-Cas9 system.

摘要: 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 promising drug targets to chemotherapy against due their essential roles absence from hosts. The objectives present study investigate functions six protein (CDPK4, CDPK4A, CDPK5, CDPK6, CDPK8, CDPK9) assess whether they suitable for designing targets. We used CRISPR-Cas9 system disrupt CDPK genes successfully insertion DHFR* at guide RNA-targeted region endogenous loci obtained knockout (KO)-CDPK strains. biological characteristics strains evaluated plaque assays, invasion, egress, replication, virulence respectively. results indicated that there was no significant difference between KO-CDPK wild-type strain lytic cycle including replication. conclusion CDPKs not also factors mice, suggesting may participate other gondii.