Department of Molecular Protozoology, Research Institute for Microbial Diseases, Osaka University, Osaka, Japan.
Center for Infectious Disease Education and Research, Osaka University, Osaka, Japan.
Nat Commun. 2022 Oct 18;13(1):6163. doi: 10.1038/s41467-022-33804-w.
The global spread of drug resistance is a major obstacle to the treatment of Plasmodium falciparum malaria. The identification of drug-resistance genes is an essential step toward solving the problem of drug resistance. Here, we report functional screening as a new approach with which to identify drug-resistance genes in P. falciparum. Specifically, a high-coverage genomic library of a drug-resistant strain is directly generated in a drug-sensitive strain, and the resistance gene is then identified from this library using drug screening. In a pilot experiment using the strain Dd2, the known chloroquine-resistant gene pfcrt is identified using the developed approach, which proves our experimental concept. Furthermore, we identify multidrug-resistant transporter 7 (pfmdr7) as a novel candidate for a mefloquine-resistance gene from a field-isolated parasite; we suggest that its upregulation possibly confers the mefloquine resistance. These results show the usefulness of functional screening as means by which to identify drug-resistance genes.
全球耐药性的传播是治疗恶性疟原虫疟疾的主要障碍。鉴定耐药基因是解决耐药性问题的关键步骤。在这里，我们报告了一种新的功能筛选方法，用于鉴定恶性疟原虫中的耐药基因。具体来说，在敏感株中直接生成耐药株的高覆盖率基因组文库，然后通过药物筛选从该文库中鉴定耐药基因。在使用 Dd2 株的初步实验中，我们使用开发的方法鉴定了已知的氯喹耐药基因 pfcrt，这证明了我们的实验概念。此外，我们还从一株分离自现场的寄生虫中鉴定出多药耐药转运蛋白 7（pfmdr7）作为氯胍耐药基因的新候选基因；我们认为其上调可能赋予了氯胍耐药性。这些结果表明功能筛选在鉴定耐药基因方面是有用的。
