Approaches for genetic manipulation of Naegleria fowleri

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Project Summary The goal of this proposal is to develop molecular tools and approaches for the study of the pathogenic free-living amoeba Naegleria fowleri. These currently do not exist. This amoeba is the causative agent of primary amebic meningoencephalitis (PAM), an almost invariable lethal brain infection. The lack of useful drugs for the treatment of this infection emphasizes the need to develop better therapeutic agents for amoebae infections. The lack of understanding of gene function in the parasite remains a major hurdle in target-based drug development campaigns. Here, we propose to address this problem by developing approaches for gene silencing and ablation by RNAi and CRISP/Cas9-mediate gene editing and transgene expression. These approaches will serve as an important first step in understanding both gene function and essentiality, key to early-stage drug discovery efforts.