使用CRISPR-Cas9系统对Toxoplasma gondii6种真核细胞翻译启动因子的功能性表征
Yong-Jie Kou1,2, Jin Gao1,2, Rui Li2
1Laboratory of Parasitic Diseases, College of Veterinary Medicine, Shanxi Agricultural University, Taigu, Jinzhong 030801, China.
International journal of molecular sciences
|July 27, 2024
概括
在Toxoplasma gondii中删除真核转化启动因子 (eIF) 揭示了TGGT1_286090对于布拉迪佐इट分化至关重要. 这一因素是治疗毒素菌症的潜在药物标.
科学领域:
- 分子生物学分子生物学
- 寄生虫学的寄生虫学
- 细胞生物学 细胞生物学
背景情况:
- 细胞转化启动因子 (eIFs) 调节蛋白质合成.
- 像Toxoplasma gondii这样的虫寄生虫依赖于精确的翻译控制.
- 了解eIF的功能是控制寄生虫感染的关键.
研究的目的:
- 调查六个eIFs在Toxoplasma gondii中的作用.
- 为了确定这些eIFs的亚细胞局部.
- 评估eIF基因删除对寄生虫适应性和差异化的影响.
主要方法:
- 在CRISPR-Cas9基因编辑中,删除了6个eIF基因.
- 对于蛋白质定位的C端内源标记和免疫光学.
- 在小鼠体内测试 (斑块,入侵,生长,退出) 和体内毒性测试.
- 分析塔基佐伊特与布拉迪佐伊特的分化.
主要成果:
- 四个eIFs定位在细胞质上,一个在细胞质上,一个在基底复合体/细胞质上.
- 删除五个eIF基因对体外生长,入侵或小鼠毒性没有显著影响.
- 删除TGGT1_286090 (RHΔ286090) 略有减少了入侵和毒性.
- TGGT1_286090 删除显著损害了塔基佐酸到布拉迪佐酸的分化.
结论:
- TGGT1_286090对于Toxoplasma gondii bradyzoite的分化至关重要.
- 在经过测试的条件下,其他被调查的eIF在RH菌株中扮演着不重要的角色.
- TGGT1_286090代表了一种潜在的治疗点,用于开发抗毒素菌的药物.
- 针对TGGT1_286090可能会导致减弱的疫苗.
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