Co-Transfection of Messenger RNA and siRNA as a Method to Study the Efficiency of siRNA

Abstract

The definition of an optimal siRNA results from the in vitro testing of several siRNA designed to specifically target a gene. Usually, such in vitro tests consist in the transfection of the several siRNA duplexes in a cell expressing stably the gene of interest. When a siRNA specific for a mRNA coding toxic proteins (certain transcription factors, transporters, toxins, cell cycle controlling proteins, etc.) must be tested, the generation of a target cell is difficult. Here we report a quick method to test the efficiency of a siRNA through its co-transfection with the targeted mRNA. This technique can be used as a fast method to test siRNA even when they target genes that cannot be stably expressed in the cells of interest.

^*Both authors contributed equally to the work.

This work was supported by “Förderprogramm Biotechnologie,” a grant from the government of Baden Württemberg, Germany and by a “IZKF-Verbundprojekt,” a grant from the University of Tübingen. JP is supported by the DFG: Graduiertenkolleg “Infektionsbiologie” in Tübingen and JPC is supported by a “Fortüne” grant from the University of Tübingen.

Keywords:

• siRNA
• Messenger RNA
• Transfection
• Electroporation
• Toxic gene

Notes

^*Both authors contributed equally to the work.

This work was supported by “Förderprogramm Biotechnologie,” a grant from the government of Baden Württemberg, Germany and by a “IZKF-Verbundprojekt,” a grant from the University of Tübingen. JP is supported by the DFG: Graduiertenkolleg “Infektionsbiologie” in Tübingen and JPC is supported by a “Fortüne” grant from the University of Tübingen.