Definition of constitutive and stage-enriched promoters in the rodent malaria parasite, Plasmodium yoelii.
Definition of constitutive and stage-enriched promoters in the rodent malaria parasite, Plasmodium yoelii.
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DOI：
10.1186/s12936-020-03498-w
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发表时间：
2020-11-23
期刊：
影响因子：
3
通讯作者：
Lindner SE
中科院分区：
文献类型：
--
作者：
Bowman LM;Finger LE;Hart KJ;Lindner SE
明确定义的启动子是所有生物体遗传研究的基本要素，可以实现内源基因的受控表达、转基因表达和基因编辑。尽管如此，对于啮齿动物传染性疟疾寄生虫，仍缺乏确定的启动子。对于约氏疟原虫来说尤其如此，它经常被用来研究疟疾感染的蚊子和肝脏阶段，以及宿主对感染的免疫反应。这里从寄生虫的整个生命周期中选择了六个启动子（clag-a、动力蛋白重链δ、lap4、trap、uis4、lisp2），这些启动子在文献中被引用为以特定阶段的方式控制其基因。还确定了赋予强表达水平的组成型 pybip 启动子的最小启动子长度，这对于报告基因和基因编辑酶的表达很有用。相反，据观察，这些启动子赋予阶段富集的基因控制，因为一些寄生虫也在其他阶段有效地使用这些启动子。因此，当单独使用时，这些启动子可能会使启动子交换、阶段性重组或基因编辑实验获得的结果的解释复杂化。这些数据共同表明，实现特定阶段的效果（例如基因编辑）可能最好使用双组件系统来实现，该系统具有仅在预期生命周期阶段重叠的独立启动子活性。
Well-defined promoters are essential elements for genetic studies in all organisms, and enable controlled expression of endogenous genes, transgene expression, and gene editing. Despite this, there is a paucity of defined promoters for the rodent-infectious malaria parasites. This is especially true for Plasmodium yoelii, which is often used to study the mosquito and liver stages of malarial infection, as well as host immune responses to infection. Here six promoters were selected from across the parasite’s life cycle (clag-a, dynein heavy chain delta, lap4, trap, uis4, lisp2) that have been invoked in the literature as controlling their genes in a stage-specific manner. A minimal promoter length for the constitutive pybip promoter that confers strong expression levels was also determined, which is useful for expression of reporters and gene editing enzymes. Instead, it was observed that these promoters confer stage-enriched gene control, as some parasites also effectively use these promoters in other stages. Thus, when used alone, these promoters could complicate the interpretation of results obtained from promoter swaps, stage-targeted recombination, or gene editing experiments. Together these data indicate that achieving stage-specific effects, such as gene editing, is likely best done using a two-component system with independent promoter activities overlapping only in the intended life cycle stage.
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