Abstract
The rodent malaria parasite Plasmodium chabaudi chabaudi shares many features with human malaria species, including P. falciparum, and is the in vivo model of choice for many aspects of malaria research in the mammalian host, from sequestration of parasitized erythrocytes, to antigenic variation and host immunity and immunopathology. This protocol describes an optimized method for the transformation of mature blood-stage P.c. chabaudi and a description of a vector that targets efficient, single crossover integration into the P.c. chabaudi genome. Transformed lines are reproducibly generated and selected within 14–20 d, and show stable long-term protein expression even in the absence of drug selection. This protocol, therefore, provides the scientific community with a robust and reproducible method to generate transformed P.c. chabaudi parasites expressing fluorescent, bioluminescent and model antigens that can be used in vivo to dissect many of the fundamental principles of malaria infection.
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Acknowledgements
We thank B. Franke-Fayard (Leiden University Medical Centre, The Netherlands) for the kind gift of plasmid pL0017. This work was supported by the Medical Research Council (MRC; reference U117584248) and the Wellcome Trust (048684). P.J.S. is the recipient of a fellowship from the Leverhulme Trust, and J. Lawton is funded by an MRC PhD studentship.
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P.J.S., D.C., W.J., J. Langhorne and J.T. designed the experiments and wrote the manuscript. P.J.S., D.C., W.J. and J. Lawton performed the parasite purification, transfection optimization and parasite visualization experiments. J.T. designed and generated transfection plasmids.
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Spence, P., Cunningham, D., Jarra, W. et al. Transformation of the rodent malaria parasite Plasmodium chabaudi. Nat Protoc 6, 553–561 (2011). https://doi.org/10.1038/nprot.2011.313
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DOI: https://doi.org/10.1038/nprot.2011.313
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