A quantitative analysis of Plasmodium falciparum transfection using DNA-loaded erythrocytes

Genetic modification of Plasmodium falciparum is a key molecular tool for the investigation of the biology and pathogenesis of this important human pathogen. The most effective means to introduce exogenous DNA into P. falciparum is via passive uptake following invasion into a DNA-loaded erythrocyte. Using bioluminescence as a tool to quantify transfection efficiency, parameters previously judged empirically to enhance transfection efficiency were subjected to a quantitative analysis. This report supports roles for fresh erythrocytes and growth medium supplemented with human serum in enhancing transfection efficiency. Critically, a proposed enhancement to transfection efficiency through continued feeding with DNA-loaded erythrocytes is not borne out in this study, and actually appears to be detrimental.

240Highlights► A quantitative assessment of key parameters for transfection of Plasmodium falciparum using DNA-loaded erythrocytes. ► Fresh erythrocytes and human serum-supplemented growth medium enhance transfection efficiencies. ► Continued feeding with DNA-loaded erythrocytes is detrimental to transfection efficiency.