Transfection of Plasmodium falciparum
Genetic manipulation of Plasmodium falciparum remains very challenging, mainly due to the parasite genome’s high A/T-richness and low transfection efficiency. This chapter includes methods for generating transient and stable transfections by electroporation, allelic replacement with tagged genes, gene deletion, and the analysis of all the above.
Key wordsPlasmodium falciparum Transfection Electroporation Vectors Positive selectable markers Negative selectable markers Knockout Allelic replacement
Methods like the ones described above are based on incremental findings and observations made by many researchers and only a few of them can be accredited for their contributions. Many members of our laboratories past and present have refined the methods that we described above. We are very grateful to all their contributions, especially to those of Alan Cowman, Brendan Crabb, Tania de Koning-Ward, Jenny Thompson, Tony Triglia, Rebecca O’Donnell, Matthew O’Neill, and Ping Cannon. We thank the Red Cross for their continuous supply of Blood and human serum. Our laboratories are supported by the Australian National Health and Medical Research Council and the Australian Research Council.