Abstract
Synchronisation of Plasmodium cultures is essential to investigate the complexities of time-dependent events associated with the asexual blood stage of the malaria parasite life cycle. Here we describe a procedure using ML10, a highly specific inhibitor of the parasite cyclic GMP-dependent protein kinase (PKG), to attain high synchronicity of Plasmodium falciparum and P. knowlesi asexual blood-stage cultures and to obtain high levels of arrested mature schizonts as well as viable released merozoites. Additionally, we describe how to use ML10 to improve the transfection efficiency of P. falciparum parasites and also how to derive the half maximal effective concentration (EC50) of ML10 in other P. falciparum laboratory lines and clinical isolates.
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Acknowledgments
We thank Gisela Henriques, Franziska Mohring, Christian Flueck, Stefanie Nofal, and Avnish Patel for their expert advice, Melissa Hart for supplying Plasmodium knowlesi parasites and James Thomas for contributing toward the original published research. Finally, we thank Simon Osborne at LifeArc for supplying ML10.
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Ressurreição, M., Moon, R.W., Baker, D.A., van Ooij, C. (2022). Synchronisation of Plasmodium falciparum and P. knowlesi In Vitro Cultures Using a Highly Specific Protein Kinase Inhibitor. In: Jensen, A.T.R., Hviid, L. (eds) Malaria Immunology. Methods in Molecular Biology, vol 2470. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-2189-9_10
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DOI: https://doi.org/10.1007/978-1-0716-2189-9_10
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