TransIT-TKO® Transfection Reagent

Figure 1. Efficient Target Gene Knockdown in Selected Cell Lines Using TransIT-TKO® Reagent. Reporter plasmids expressing both firefly and sea pansy luciferases were co-transfected into the indicated cell lines using TransIT® Plasmid Transfection Reagents. Targeted knockdown was achieved by transfection of an anti-firefly luciferase siRNA using the TransIT-TKO® Reagent. Twenty-four hours later, firefly luciferase expression was normalized to sea pansy luciferase expression and compared to the reagent alone control.

Figure 2. Efficient Knockdown of Endogenous Genes in Primary Hepatocytes Using TransIT-TKO® Reagent. Primary mouse hepatocytes were transfected with the indicated siRNAs or a non-targeting control siRNA using the TransIT-TKO® Reagent. Twenty-four hours post-transfection, the amount of each mRNA was measured relative to GAPDH mRNA levels using qRT-PCR and then scaled to the expression level of the specific target mRNA in the cells alone (untreated) controls.

Figure 3. Efficient Knockdown of Luciferase Expression Using TransIT-TKO® Reagent to Deliver Nanomolar Amounts of siRNA. Reporter plasmids expressing firefly and sea pansy luciferase were transfected into COS-7 cells using the TransIT®-LT1 Reagent. After four hours, various concentrations of anti-firefly luciferase siRNA (0, 0.05, 0.25, 0.75, 1, 5, 10, and 25 nM) were complexed with the TransIT-TKO® Reagent and transfected into COS-7 cells in their complete media. Twenty-four hours post-transfection, firefly luciferase expression was normalized to sea pansy luciferase expression and compared to the reagent alone control.

Figure 4. Knockdown of Endogenous Genes Using TransIT-TKO® Reagent. Various cells were transfected with siRNAs targeting the indicated genes using the TransIT-TKO® Reagent, and the knockdown percentage was determined using quantitative RT-PCR.

Figure 5. Visualization of Fluorescently Labeled siRNA. siRNA was fluorescently labeled with the Label IT® siRNA Tracker Fluorescein Kit and transfected into HeLa cells using the TransIT-TKO® Reagent. At 24 hours, cells were fixed, counterstained, and analyzed by confocal microscopy to visualize the siRNA (green), nuclei (blue), and actin (red).