CRISPR/Cas9-engineered inducible gametocyte producer lines as a valuable tool for Plasmodium falciparum malaria transmission research

Sylwia D. Boltryk, Armin Passecker, Arne Alder, Eilidh Carrington, Marga Vegte-Bolmer, Geert-Jan Gemert, Alex Starre, Hans-Peter Beck, Robert W. Sauerwein, Taco W. A. Kooij, Nicolas M. B. Brancucci, Nicholas I. Proellochs, Tim-Wolf Gilberger and Till S. Voss ()
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Sylwia D. Boltryk: Swiss Tropical and Public Health Institute
Armin Passecker: Swiss Tropical and Public Health Institute
Arne Alder: Centre for Structural Systems Biology
Eilidh Carrington: Swiss Tropical and Public Health Institute
Marga Vegte-Bolmer: Radboud University Medical Center
Geert-Jan Gemert: Radboud University Medical Center
Alex Starre: Radboud University Medical Center
Hans-Peter Beck: Swiss Tropical and Public Health Institute
Robert W. Sauerwein: Radboud University Medical Center
Taco W. A. Kooij: Radboud University Medical Center
Nicolas M. B. Brancucci: Swiss Tropical and Public Health Institute
Nicholas I. Proellochs: Radboud University Medical Center
Tim-Wolf Gilberger: Centre for Structural Systems Biology
Till S. Voss: Swiss Tropical and Public Health Institute

Nature Communications, 2021, vol. 12, issue 1, 1-16

Abstract: Abstract The malaria parasite Plasmodium falciparum replicates inside erythrocytes in the blood of infected humans. During each replication cycle, a small proportion of parasites commits to sexual development and differentiates into gametocytes, which are essential for parasite transmission via the mosquito vector. Detailed molecular investigation of gametocyte biology and transmission has been hampered by difficulties in generating large numbers of these highly specialised cells. Here, we engineer P. falciparum NF54 inducible gametocyte producer (iGP) lines for the routine mass production of synchronous gametocytes via conditional overexpression of the sexual commitment factor GDV1. NF54/iGP lines consistently achieve sexual commitment rates of 75% and produce viable gametocytes that are transmissible by mosquitoes. We also demonstrate that further genetic engineering of NF54/iGP parasites is a valuable tool for the targeted exploration of gametocyte biology. In summary, we believe the iGP approach developed here will greatly expedite basic and applied malaria transmission stage research.

Date: 2021
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DOI: 10.1038/s41467-021-24954-4

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