Generation of Transmission-Competent Human Malaria Parasites with Chromosomally-Integrated Fluorescent Reporters.

TitleGeneration of Transmission-Competent Human Malaria Parasites with Chromosomally-Integrated Fluorescent Reporters.
Publication TypeJournal Article
Year of Publication2019
AuthorsMcLean KJarrod, Straimer J, Hopp CS, Vega-Rodriguez J, Small-Saunders JL, Kanatani S, Tripathi A, Mlambo G, Dumoulin PC, Harris CT, Tong X, Shears MJ, Ankarklev J, Kafsack BFC, Fidock DA, Sinnis P
JournalSci Rep
Date Published2019 09 11
KeywordsAnimals, Culicidae, Flow Cytometry, Genetic Engineering, Green Fluorescent Proteins, Humans, Life Cycle Stages, Luminescent Proteins, Malaria, Falciparum, Microscopy, Fluorescence, Parasites, Phenotype, Plasmodium falciparum, Protozoan Proteins

Malaria parasites have a complex life cycle that includes specialized stages for transmission between their mosquito and human hosts. These stages are an understudied part of the lifecycle yet targeting them is an essential component of the effort to shrink the malaria map. The human parasite Plasmodium falciparum is responsible for the majority of deaths due to malaria. Our goal was to generate transgenic P. falciparum lines that could complete the lifecycle and produce fluorescent transmission stages for more in-depth and high-throughput studies. Using zinc-finger nuclease technology to engineer an integration site, we generated three transgenic P. falciparum lines in which tdtomato or gfp were stably integrated into the genome. Expression was driven by either stage-specific peg4 and csp promoters or the constitutive ef1a promoter. Phenotypic characterization of these lines demonstrates that they complete the life cycle with high infection rates and give rise to fluorescent mosquito stages. The transmission stages are sufficiently bright for intra-vital imaging, flow cytometry and scalable screening of chemical inhibitors and inhibitory antibodies.

Alternate JournalSci Rep
PubMed ID31511546
PubMed Central IDPMC6739413
Grant ListF31 AI136405 / AI / NIAID NIH HHS / United States
R01 AI132359 / AI / NIAID NIH HHS / United States
T32 AI100852 / AI / NIAID NIH HHS / United States

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