Nature Microbiology ( IF 20.5 ) Pub Date : 2024-07-08 , DOI: 10.1038/s41564-024-01754-2
Christopher J Giuliano 1, 2 , Kenneth J Wei 1, 2 , Faye M Harling 1 , Benjamin S Waldman 1, 2 , Madeline A Farringer 3, 4 , Elizabeth A Boydston 1 , Tammy C T Lan 1 , Raina W Thomas 1, 2 , Alice L Herneisen 1, 2 , Allen G Sanderlin 2 , Isabelle Coppens 5 , Jeffrey D Dvorin 3, 6 , Sebastian Lourido 1, 2
Examining host–pathogen interactions in animals can capture aspects of infection that are obscured in cell culture. Using CRISPR-based screens, we functionally profile the entire genome of the apicomplexan parasite Toxoplasma gondii during murine infection. Barcoded gRNAs enabled bottleneck detection and mapping of population structures within parasite lineages. Over 300 genes with previously unknown roles in infection were found to modulate parasite fitness in mice. Candidates span multiple axes of host–parasite interaction. Rhoptry Apical Surface Protein 1 was characterized as a mediator of host-cell tropism that facilitates repeated invasion attempts. GTP cyclohydrolase I was also required for fitness in mice and druggable through a repurposed compound, 2,4-diamino-6-hydroxypyrimidine. This compound synergized with pyrimethamine against T. gondii and malaria-causing Plasmodium falciparum parasites. This work represents a complete survey of an apicomplexan genome during infection of an animal host and points to novel interfaces of host–parasite interaction.
中文翻译:
急性小鼠感染期间弓形虫基因组基于 CRISPR 的功能分析
检查动物体内的宿主-病原体相互作用可以捕获细胞培养中被掩盖的感染方面。使用基于 CRISPR 的筛选,我们在小鼠感染期间对顶复门寄生虫弓形虫的整个基因组进行了功能分析。条形码 gRNA 能够对寄生虫谱系中的种群结构进行瓶颈检测和作图。发现 300 多个以前在感染中作用未知的基因可调节小鼠的寄生虫适应性。候选药物跨越宿主-寄生虫相互作用的多个轴。Rhoptry 顶端表面蛋白 1 被表征为宿主细胞嗜性的介质,促进反复侵袭尝试。GTP 环水解酶 I 也是小鼠健身所必需的,并且可通过重新利用的化合物 2,4-二氨基-6-羟基嘧啶成药。该化合物与乙胺嘧啶协同作用,对抗刚地弓形虫和引起疟疾的恶性疟原虫寄生虫。这项工作代表了动物宿主感染过程中顶复体基因组的完整调查,并指出了宿主-寄生虫相互作用的新界面。