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

Malar J. 2012 Jun 21;11:210. doi: 10.1186/1475-2875-11-210.

Abstract

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 9.59 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.

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.

Publication types

  • Comparative Study
  • Evaluation Study
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Electroporation / methods*
  • Genes, Reporter
  • Luciferases / analysis
  • Luciferases / genetics
  • Molecular Biology / methods*
  • Parasitology / methods*
  • Plasmodium falciparum / genetics*
  • Transfection / methods*

Substances

  • Luciferases