Thomas Phaedra, Sedillo Jennifer, Oberstaller Jenna, Li Suzanne, Zhang Min, Singh Naresh, Wang Chengqi C Q, Udenze Kenneth, Jiang Rays H Y, Adams John H
Center for Global Health and Infectious Diseases Research, Department of Global Health, University of South Florida, Tampa, Florida, USA.
mSphere. 2016 Oct 26;1(5). doi: 10.1128/mSphere.00273-16. eCollection 2016 Sep-Oct.
Malaria remains one of the most devastating parasitic diseases worldwide, with 90% of the malaria deaths in Africa in 2013 attributable to . The clinical symptoms of malaria include cycles of fever, corresponding to parasite rupture from red blood cells every 48 h. Parasite pathways involved in the parasite's ability to survive the host fever response, and indeed, the functions of ~40% of genes as a whole, are still largely unknown. Here, we evaluated the potential of scalable forward-genetic screening methods to identify genes involved in the host fever response. We performed a phenotypic screen for genes linked to the parasite response to febrile temperatures by utilizing a selection of single-disruption mutants generated via random transposon mutagenesis in a previous study. We identified several mutants presenting significant phenotypes in febrile response screens compared to the wild type, indicating possible roles for the disrupted genes in this process. We present these initial studies as proof that forward genetics can be used to gain insight into critical factors associated with parasite biology. Though the genome sequence has been available for many years, ~40% of its genes do not have informative annotations, as they show no detectable homology to those of studied organisms. More still have not been evaluated via genetic methods. Scalable forward-genetic approaches that allow interrogation of gene function without any pre-existing knowledge are needed to hasten understanding of parasite biology, which will expedite the identification of drug targets and the development of future interventions in the face of spreading resistance to existing frontline drugs. In this work, we describe a new approach to pursue forward-genetic phenotypic screens for to identify factors associated with virulence. Future large-scale phenotypic screens developed to probe other such interesting phenomena, when considered in parallel, will prove a powerful tool for functional annotation of the genome, where so much remains undiscovered.
疟疾仍然是全球最具毁灭性的寄生虫病之一，2013年非洲90%的疟疾死亡病例都可归因于此。疟疾的临床症状包括周期性发热，这与寄生虫每48小时从红细胞中破裂有关。寄生虫在宿主发热反应中存活的能力所涉及的寄生虫途径，实际上，总体上约40%的基因功能仍 largely unknown。在这里，我们评估了可扩展的正向遗传学筛选方法识别参与宿主发热反应基因的潜力。我们通过利用在先前研究中通过随机转座子诱变产生的单破坏突变体选择，对与寄生虫对发热温度反应相关的基因进行了表型筛选。与野生型相比，我们在发热反应筛选中鉴定出几个呈现显著表型的突变体，表明被破坏的基因在此过程中可能发挥的作用。我们展示这些初步研究作为证据，证明正向遗传学可用于深入了解与寄生虫生物学相关的关键因素。尽管该基因组序列已经可用多年，但其约40%的基因没有信息丰富的注释，因为它们与已研究生物的基因没有可检测到的同源性。更多的基因仍未通过遗传方法进行评估。需要可扩展的正向遗传学方法，在没有任何先验知识的情况下对基因功能进行研究，以加速对寄生虫生物学的理解，这将加快药物靶点的识别以及面对对现有一线药物耐药性不断蔓延时未来干预措施的开发。在这项工作中，我们描述了一种新方法，用于对该寄生虫进行正向遗传学表型筛选，以识别与毒力相关的因素。未来开发的用于探索其他此类有趣现象的大规模表型筛选，若并行考虑，将证明是该寄生虫基因组功能注释的有力工具，因为仍有许多未知之处。
