Characterization of functions in parasite growth and virulence of four Toxoplasma gondii genes involved in lipid synthesis by CRISPR-Cas9 system
Characterization of functions in parasite growth and virulence of four Toxoplasma gondii genes involved in lipid synthesis by CRISPR-Cas9 system
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DOI：
10.1007/s00436-021-07308-3
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发表时间：
2021-09
影响因子：
2
通讯作者：
Jie Liu;Ting-Ting Li-Ting;Qin-Li Liang;H. Elsheikha;Dan Zhao;Zhi-Wei Zhang;Xiao-pei Xu;Xing-Quan Zhu;Meng Wang
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文献类型：
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作者：
Jie Liu;Ting-Ting Li-Ting;Qin-Li Liang;H. Elsheikha;Dan Zhao;Zhi-Wei Zhang;Xiao-pei Xu;Xing-Quan Zhu;Meng Wang
脂肪酸摄取对细胞内寄生虫刚地弓形虫的生存和生长极为重要。本研究利用CRISPR-Cas9基因编辑技术，研究了甘油-3-磷酸脱氢酶（G3PDH）、丙二酰辅酶a -酰基载体蛋白转酰基酶（FabD）、酰基- acp硫酯酶（TE）和二酰基甘油酰基转移酶（DGAT）四种脂质合成酶在I型RH和II型Prugniaud菌株ofT毒力和感染性中的作用。刚。速殖子期免疫荧光分析显示，FabD蛋白位于顶质体；然而，其他三种蛋白的表达水平未检测到。与野生型（WT）菌株相比，RHΔG3PDH、RHΔTE和RHΔDGATin的体外生长和体内毒力无显著差异。但是，RHΔFabDexhibited与WT菌株相比，生长速率明显降低。该基因的缺失降低了II型Pru菌株的毒力，减少了体内囊肿的形成。这些数据提高了我们对脂质合成酶在脂肪肝发病机制中的作用的理解。刚。
Fatty acid uptake is extremely important for the survival and growth of the intracellular parasiteToxoplasma 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 ofT. 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 RHΔDGATin vitro and their virulence in vivo were not significantly different. However, RHΔFabDexhibited a significantly reduced growth rate, compared with the WT strain. The deletion ofFabDattenuated 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 ofT. gondii.