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

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. A time-course of luciferase reporter expression is used to provide an objective quantitative analysis of the a...

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Bibliographic Details
Published in:Malaria journal 2012-06, Vol.11 (1), p.210-210, Article 210
Main Authors: Hasenkamp, Sandra, Russell, Karen T, Horrocks, Paul
Format: Article
Language:English
Subjects:
Comparative analysis
DNA
Electroporation
Electroporation - methods
Erythrocytes
Genes, Reporter
Genetic aspects
Health aspects
Luciferase
Luciferases - analysis
Luciferases - genetics
Malaria
Methodology
Molecular Biology - methods
Parasites
Parasitology - methods
Physiological aspects
Plasmids
Plasmodium falciparum
Plasmodium falciparum - genetics
Studies
Transfection - methods
Transfection efficiency
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Summary: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. 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. Preloading of erythrocytes shows a mean efficiency of 9.59 x 10(-6), some 5-180 fold more efficient than matched experiments utilizing the "double-tap" and direct electroporation of ring stage infected erythrocytes alone, respectively. 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.
ISSN:1475-2875
1475-2875
DOI:10.1186/1475-2875-11-210