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

Scientific reports (2019-09-13)
Kyle Jarrod McLean, Judith Straimer, Christine S Hopp, Joel Vega-Rodriguez, Jennifer L Small-Saunders, Sachie Kanatani, Abhai Tripathi, Godfree Mlambo, Peter C Dumoulin, Chantal T Harris, Xinran Tong, Melanie J Shears, Johan Ankarklev, Björn F C Kafsack, David A Fidock, Photini Sinnis
ABSTRACT

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.

MATERIALS
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Sigma-Aldrich
Xanthurenic acid, 96%