Functional Characterization of Eight Zinc Finger Motif-Containing Proteins in Toxoplasma gondii Type I RH Strain Using the CRISPR-Cas9 System

文献类型： 外文期刊

第一作者： Gao, Jin

作者： Gao, Jin;Wu, Xiao-Jing;Zheng, Xiao-Nan;Kou, Yong-Jie;Zhu, Xing-Quan;Gao, Jin;Wu, Xiao-Jing;Li, Ting-Ting;Kou, Yong-Jie;Wang, Xin-Cheng;Wang, Meng;Zhu, Xing-Quan

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关键词： Toxoplasma gondii; toxoplasmosis; zinc finger protein; CRISPR-Cas9; replication

期刊名称：PATHOGENS （ 影响因子：3.7； 五年影响因子：3.7 ）

ISSN：

年卷期： 2023 年 12 卷 10 期

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收录情况： SCI

摘要： The Zinc finger protein (ZFP) family is widely distributed in eukaryotes and interacts with DNA, RNA, and various proteins to participate in many molecular processes. In the present study, the biological functions of eight ZFP genes in the lytic cycle and the pathogenicity of Toxoplasma gondii were examined using the CRISPR-Cas9 system. Immunofluorescence showed that four ZFPs (RH248270-HA, RH255310-HA, RH309200-HA, and RH236640-HA) were localized in the cytoplasm, and one ZFP (RH273150-HA) was located in the nucleus, while the expression level of RH285190-HA, RH260870-HA, and RH248450-HA was undetectable. No significant differences were detected between seven RH Delta zfp strains (RH Delta 285190, RH Delta 248270, RH Delta 260870, RH Delta 255310, RH Delta 309200, RH Delta 248450, and RH Delta 236640) and the wild-type (WT) strain in the T. gondii lytic cycle, including plaque formation, invasion, intracellular replication, and egress, as well as in vitro virulence (p > 0.05). However, the RH Delta 273150 strain exhibited significantly lower replication efficiency compared to the other seven RH Delta zfp strains and the WT strain, while in vivo virulence in mice was not significantly affected. Comparative expression analysis of the eight zfp genes indicates that certain genes may have essential functions in the sexual reproductive stage of T. gondii. Taken together, these findings expand our current understanding of the roles of ZFPs in T. gondii.

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