Department of Global Health, College of Public Health, University of South Florida, Tampa, Florida, United States of America.
PLoS One. 2010 Oct 11;5(10):e13282. doi: 10.1371/journal.pone.0013282.
A majority of the Plasmodium falciparum genome codes for genes with unknown functions, which presents a major challenge to understanding the parasite's biology. Large-scale functional analysis of the parasite genome is essential to pave the way for novel therapeutic intervention strategies against the disease and yet difficulties in genetic manipulation of this deadly human malaria parasite have been a major hindrance for functional analysis of its genome. Here, we used a forward functional genomic approach to study P. falciparum and identify genes important for optimal parasite development in the disease-causing, intraerythrocytic stages. We analyzed 123 piggyBac insertion mutants of P. falciparum for proliferation efficiency in the intraerythrocytic stages, in vitro. Almost 50% of the analyzed mutants showed significant reduction in proliferation efficiency, with 20% displaying severe defects. Functional categorization of genes in the severely attenuated mutants revealed significant enrichment for RNA binding proteins, suggesting the significance of post-transcriptional gene regulation in parasite development and emphasizing its importance as an antimalarial target. This study demonstrates the feasibility of much needed forward genetics approaches for P. falciparum to better characterize its genome and accelerate drug and vaccine development.
疟原虫基因组的大部分编码具有未知功能的基因，这给理解寄生虫的生物学带来了重大挑战。对寄生虫基因组进行大规模功能分析对于开辟针对该疾病的新型治疗干预策略至关重要，但对这种致命的人类疟疾寄生虫进行遗传操作的困难一直是其基因组功能分析的主要障碍。在这里，我们使用正向功能基因组学方法来研究疟原虫，并确定在致病的红细胞内阶段对寄生虫最佳发育至关重要的基因。我们分析了 123 个疟原虫 piggyBac 插入突变体在红细胞内阶段的增殖效率，在体外。分析的突变体中几乎有 50%的增殖效率显著降低，其中 20%显示出严重的缺陷。严重衰减突变体中基因的功能分类显示出 RNA 结合蛋白的显著富集，表明转录后基因调控在寄生虫发育中的重要性，并强调了其作为抗疟靶点的重要性。这项研究证明了针对疟原虫进行正向遗传学研究的必要性，以更好地描述其基因组并加速药物和疫苗的开发。
