Comparison of the absolute and relative efficiencies of electroporation-based transfection protocols for Plasmodium falciparum

Background: Several electroporation protocols exist to transfect exogenous DNA into Plasmodium falciparum. To date, however, only a subjective analysis of their relative efficiencies has been reported. Methods: A time-course of luciferase reporter expression is used to provide an objective quantitative analysis of the absolute efficiency of three electroporation techniques; direct electroporation of ring stage infected erythrocytes, preloading of erythrocytes and a novel double-tap protocol that combines both approaches. Results: Preloading of erythrocytes shows a mean efficiency of 9.59x10(-6), some 5-180 fold more efficient than matched experiments utilizing the double-tap and direct electroporation of ring stage infected erythrocytes alone, respectively. Conclusion: Evidence presented here provides the first quantitative assessment of both the absolute and relative efficiencies of a key molecular tool used to study the biology and pathogenesis of this important human pathogen.