Description
The GeneSilencer^® PCR Kits rely on the PCR technology to enable the production of sequence specific siRNAs.  The desired siRNA expression cassettes are generated by a one step PCR, and are then transfected into cells. Subsequently, the cassettes are transcribed to produce sense and anti-sense siRNAs, which anneal in situ to form the duplex siRNAs that are effective for gene silencing.

The kits contain DNA templates with the U6 or the H1 RNA polymerase III promoters, allowing optimal expression in a wide variety of cell types.  Additionally, these kits are available with a GFP reporter gene cassette to allow easy determination of PCR fragment’s transfection efficiency.  .

Figure 1: Co-transfection Silencing with siRNA PCR Fragments.

Figure 1 legend: Four different siRNAs against the Luciferase gene (H1-Luc, U6-Luc, H1-GFP-Luc, U6-GFP-Luc) were produced using the different primer pairs and templates enclosed in the kit.  Synthetic Luciferase siRNA was obtained from Dharmacon, Inc. (Lafayette, CO).  Varying amounts of each siRNA were co-transfected into COS cells with 0.5µg gWiz-Luciferase plasmid using the GeneSilencer^® siRNA Transfection Reagent.  Luciferase expression levels were measured relative to background and normalized to reflect total amount of Luciferase per mg of total protein per well.  Percent silencing levels relative to gWiz-Luciferase-only expression are shown

† Custom primers are not included in kit and must be purchased seperately.