A versatile Plasmodium falciparum reporter line expressing NanoLuc enables highly sensitive multi-stage drug assays.
A versatile Plasmodium falciparum reporter line expressing NanoLuc enables highly sensitive multi-stage drug assays.
复制标题
表达纳米核的多功能疟原虫记者线可实现高度敏感的多阶段药物测定。
DOI：
10.1038/s42003-023-05078-5
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发表时间：
2023-07-12
影响因子：
5.9
通讯作者：
Miyazaki, Shinya
中科院分区：
文献类型：
--
作者：
Miyazaki, Yukiko;Vos, Martijn W.;Geurten, Fiona J. A.;Bigeard, Pierre;Kroeze, Hans;Yoshioka, Shohei;Arisawa, Mitsuhiro;Inaoka, Daniel Ken;Soulard, Valerie;Dechering, Koen J.;Franke-Fayard, Blandine;Miyazaki, Shinya
表达转基因荧光素酶的恶性疟原虫寄生虫已广泛用于评估抗疟疾化合物。在这里，为了筛选对寄生虫的多个阶段有效的抗疟疾药物，我们生成了一种恶性疟原虫报告寄生虫，该寄生虫在其整个生命周期中组成型表达纳米荧光素酶（NanoLuc）。表达 NanoLuc 的恶性疟原虫报告寄生虫在无性血液、配子体、蚊子和肝脏阶段显示定量的 NanoLuc 信号。我们还建立了测定系统来评估化合物在无性血液、配子体和肝脏阶段的抗疟活性，然后确定几种抗疟化合物的50%抑制浓度（IC50）值。通过开发这种强大的高通量筛选系统，我们确定了一种可以杀死无性血期寄生虫的抗疟化合物。我们的研究强调了 NanoLuc 报告基因系的实用性，它可以通过在多个阶段改进针对人类疟疾寄生虫的化合物的筛选来推进抗疟疾药物的开发。介绍一种多功能恶性疟原虫双报告基因系，在组成型启动子下表达荧光蛋白和 NanoLuc，可用于筛选对疟原虫多个阶段有效的新型抗疟疾药物。
Transgenic luciferase-expressing Plasmodium falciparum parasites have been widely used for the evaluation of anti-malarial compounds. Here, to screen for anti-malarial drugs effective against multiple stages of the parasite, we generate a P. falciparum reporter parasite that constitutively expresses NanoLuciferase (NanoLuc) throughout its whole life cycle. The NanoLuc-expressing P. falciparum reporter parasite shows a quantitative NanoLuc signal in the asexual blood, gametocyte, mosquito, and liver stages. We also establish assay systems to evaluate the anti-malarial activity of compounds at the asexual blood, gametocyte, and liver stages, and then determine the 50% inhibitory concentration (IC50) value of several anti-malarial compounds. Through the development of this robust high-throughput screening system, we identify an anti-malarial compound that kills the asexual blood stage parasites. Our study highlights the utility of the NanoLuc reporter line, which may advance anti-malarial drug development through the improved screening of compounds targeting the human malarial parasite at multiple stages. Presentation of a versatile Plasmodium falciparum dual reporter line, expressing both a fluorescent protein and NanoLuc under a constitutive promoter, that can be used to screen for novel anti-malarial drugs effective against multiple stages of the parasite.
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影响因子：
14.9
作者：
Deitsch, KW;Driskill, CL;Wellems, TE
通讯作者：
Wellems, TE
影响因子：
5.2
作者：
D'Alessandro, Sarah;Silvestrini, Francesco;Taramelli, Donatella
通讯作者：
Taramelli, Donatella
影响因子：
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作者：
通讯作者：
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影响因子：
4.6
作者：
Marin-Mogollon C;van de Vegte-Bolmer M;van Gemert GJ;van Pul FJA;Ramesar J;Othman AS;Kroeze H;Miao J;Cui L;Williamson KC;Sauerwein RW;Janse CJ;Khan SM
通讯作者：
Khan SM
影响因子：
4.9
作者：
Lucantoni, Leonardo;Fidock, David A.;Avery, Vicky M.
通讯作者：
Avery, Vicky M.