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Characterization of functions in parasite growth and virulence of four Toxoplasma gondii genes involved in lipid synthesis by CRISPR-Cas9 system

  • Genetics, Evolution, and Phylogeny - Original Paper
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Abstract

Fatty acid uptake is extremely important for the survival and growth of the intracellular parasite Toxoplasma gondii. In this study, CRISPR-Cas9 gene editing technology was used to investigate the role of four lipid synthesis enzymes, namely, glycerol-3-phosphate dehydrogenase (G3PDH), malonyl CoA-acyl carrier protein transacylase (FabD), acyl-ACP thiolesterase (TE), and diacylglycerol acyltransferase (DGAT), in the virulence and infectivity of Type I RH and Type II Prugniaud (Pru) strains of T. gondii. Immunofluorescence analysis of the tachyzoite stage showed that FabD protein was located in the apicoplast; however, the expression level of the other three proteins was undetectable. Compared with wild-type (WT) strains, the growth of RHΔG3PDH, RHΔTE, and RHΔDGAT in vitro and their virulence in vivo were not significantly different. However, RHΔFabD exhibited a significantly reduced growth rate, compared with the WT strain. The deletion of FabD attenuated the virulence of Type II Pru strain and reduced the formation of cysts in vivo. These data improved our understanding of the role of lipid synthesis enzymes in the pathogenesis of T. gondii.

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Acknowledgements

We thank Professor Bang Shen, Huazhong Agricultural University, for providing the pSAG1-Cas9-SgUPRT and pUPRT-DHFR-D vectors.

Funding

Project support was kindly provided by the National Natural Science Foundation of China (Grant No. 32002306), the Fund for Shanxi “1331 Project,” the Agricultural Science and Technology Innovation Program (ASTIP) of China (Grant No. CAAS-ASTIP-2016-LVRI-03), the Yunnan Expert Workstation (Grant No. 202005AF150041), and the Veterinary Public Health Innovation Team of Yunnan Province (Grant No. 202105AE160014).

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Authors and Affiliations

  1. College of Animal Science and Technology, Jilin Agricultural University, Changchun, Jilin Province, 130118, People’s Republic of China

    Jie Liu & Xing-Quan Zhu

  2. State Key Laboratory of Veterinary Etiological Biology, Key Laboratory of Veterinary Parasitology of Gansu Province, Lanzhou Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Lanzhou, Gansu Province, 730046, People’s Republic of China

    Jie Liu, Ting-Ting Li, Qin-Li Liang, Dan-Yu Zhao, Zhi-Wei Zhang, Xiao-Pei Xu, Xing-Quan Zhu & Meng Wang

  3. Faculty of Medicine and Health Sciences, School of Veterinary Medicine and Science, University of Nottingham, Loughborough, LE12 5RD, UK

    Hany M. Elsheikha

  4. College of Veterinary Medicine, Shanxi Agricultural University, Taigu, Shanxi Province, 030801, People’s Republic of China

    Xing-Quan Zhu

  5. Key Laboratory of Veterinary Public Health of Higher Education of Yunnan Province, College of Veterinary Medicine, Yunnan Agricultural University, Kunming, People’s Republic of China

    Xing-Quan Zhu

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  1. Jie Liu
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Correspondence to Xing-Quan Zhu or Meng Wang.

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The present study was reviewed and approved by the Animal Ethics Committee of Lanzhou Veterinary Research Institute, Chinese Academy of Agricultural Sciences. All animals were handled strictly according to the Animal Ethics Procedures and Guidelines of the People’s Republic of China.

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Liu, J., Li, TT., Liang, QL. et al. Characterization of functions in parasite growth and virulence of four Toxoplasma gondii genes involved in lipid synthesis by CRISPR-Cas9 system. Parasitol Res 120, 3749–3759 (2021). https://doi.org/10.1007/s00436-021-07308-3

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