@article{77ea20ae0ba6414abd8b437ba72a6683,
title = "Generation of Transmission-Competent Human Malaria Parasites with Chromosomally-Integrated Fluorescent Reporters",
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.",
author = "McLean, {Kyle Jarrod} and Judith Straimer and Hopp, {Christine S.} and Joel Vega-Rodriguez and Small-Saunders, {Jennifer L.} and Sachie Kanatani and Abhai Tripathi and Godfree Mlambo and Dumoulin, {Peter C.} and Harris, {Chantal T.} and Xinran Tong and Shears, {Melanie J.} and Johan Ankarklev and Kafsack, {Bj{\"o}rn F.C.} and Fidock, {David A.} and Photini Sinnis",
note = "Funding Information: The authors would like to thank the Bloomberg Family Foundation for their generous support of the Insectary and Parasitology core facilities at the Johns Hopkins Malaria Institute. We also thank Chris Kizito for expert mosquito rearing and the Johns Hopkins School of Medicine Microscopy Facility (MicFac). This work was supported by the National Institutes of Health (R01 AI132359 to P.S.; F31 pre-doctoral fellowship F31AI136405 to C.T.H.; Columbia Integrated Training Program in Infectious Diseases Research T32 AI100852 to J.L.S.), the Bill & Melinda Gates Foundation (OPP1040399 to D.A.F.), the Johns Hopkins Malaria Research Institute (postdoctoral fellowship to C.S.H. and pre-doctoral fellowship to P.C.D.), the Swedish Research Council (J.A.) and the Swedish Society of Medicine (J.A.). Publisher Copyright: {\textcopyright} 2019, The Author(s).",
year = "2019",
month = dec,
day = "1",
doi = "10.1038/s41598-019-49348-x",
language = "English (US)",
volume = "9",
journal = "Scientific Reports",
issn = "2045-2322",
publisher = "Nature Publishing Group",
number = "1",
}