在Plasmodium falciparum中无意中介的衰变机器是低效的和不必要的
Emma McHugh1, Michaela S Bulloch1, Steven Batinovic2
1Department of Biochemistry and Pharmacology, Bio21 Molecular Science and Biotechnology Institute, The University of Melbourne , Parkville, Victoria, Australia.
mSphere
|June 27, 2023
概括
无意中介衰变 (NMD) 蛋白质对于降解错误的转录或在Plasmodium falciparum中的寄生虫生长并不重要. 这项研究还证明了在疟疾寄生虫中有效的CRISPR-Cas9基因编辑.
科学领域:
- 分子生物学分子生物学
- 遗传学 是一个遗传学.
- 寄生虫学的寄生虫学
背景情况:
- 无意中介衰变 (NMD) 是一种关键的mRNA质量控制途径,在真核生物中保存,消除具有过早终止编码的转录.
- 在甲基动物中,NMD还通过编程内置保留来调节基因表达,这种现象在Plasmodium falciparum中观察到高水平.
研究的目的:
- 调查NMD在Plasmodium falciparum中的作用,特别是检查其参与无意义转录的降解和调节内部保留的作用.
- 评估对寄生虫生长和活力的核心NMD因子PfUPF1和PfUPF2的要求.
- 建立和验证CRISPR-Cas9基因编辑在Plasmodium falciparum中的效率.
主要方法:
- 使用CRISPR-Cas9基因编辑来破坏和标记UPF1和UPF2 (PfUPF1和PfUPF2) 的Plasmodium falciparum正基因组.
- 使用免疫光显微镜将PfUPF1和PfUPF2定位在寄生虫细胞质中.
- 进行了共免疫沉试验,以分析蛋白质相互作用.
- 进行RNA测序 (RNA-seq) 来评估转录丰度,并确定NMD和内质保留的潜在目标.
主要成果:
- PfUPF1和PfUPF2被定位到细胞质点,并被证明与彼此和其他mRNA结合蛋白相互作用.
- 尽管它们的表达和相互作用,但发现PfUPF1和PfUPF2对于P. falciparum中含有无意义的转录的降解并不需要.
- 这项研究表明,P. falciparum中的大多数内质保留事件缺乏功能意义,NMD对于寄生虫生长来说是不可或缺的.
- 证明了有效的CRISPR-Cas9编辑,简化了这一具有挑战性的生物体中的基因组修改.
结论:
- 正规的无意中介的衰变途径对于降解异常转录或Plasmodium falciparum的生长并不重要.
- 在Plasmodium falciparum中编程的内置保留似乎在很大程度上是非功能性的,并且不受NMD的调节.
- 开发的CRISPR-Cas9系统为Plasmodium falciparum的基因操纵提供了一种简化方法,促进了未来的研究.
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