TransIT-TKO® Transfection Reagent

A high efficiency, low toxicity, siRNA transfection reagent for mammalian cells

Resources

Protocol (NEW! Plasmid DNA & siRNA Co-transfection)
MSDS
Frequently Asked Questions
Technical Report: High Efficiency siRNA Delivery In Vitro
Publication: Delivery of siRNAs into Mammalian Cells...
Poster: RNAi Applications in Nucleic Acid Delivery...
Product Citations
Tips From the Bench
Contact Technical Support
Request a Sample

Don't See Your Cell Type? Ask Reagent Agent

• Broad Spectrum siRNA and Plasmid DNA Delivery - Utilize one transfection reagent and protocol for a wide variety of cells.
• Low Cellular Toxicity - Maintain cell density and reduce experimental biases.
• Reproducible Results - Obtain consistent, targeted gene knockdowns from day to day.
• High Knockdown Efficiency - Achieve optimal gene silencing in a large percentage of cells to ensure experimental success.

Data

Figure 1: Efficient Target Gene Knockdown in Selected Cell Lines using TransIT-TKO Reagent.
Figure 2: Efficient Knockdown of Endogenous Genes in Primary Hepatocytes Using TransIT-TKO Reagent.
Figure 3: Efficient Knockdown of Luciferase Expression Using TransIT-TKO Reagent.
Figure 4: Knockdown of Endogenous Genes Using TransIT-TKO Reagent.
Figure 5: Visualization of Fluorescently Labeled siRNA.

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.

Back to Top

 

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.

Back to Top

 

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.

Back to Top

 

Cell Line (Source)	Endogenous Transcript	Knockdown Efficiency
BNL CL.2 (mouse liver)	MAPK1	80%
	MAPK3	83%
HeLa (human cervix)	Lamin A/C	80%
	GAPDH	80%
Hepa1c1c7 (mouse liver)	MAPK1	80%
	MAPK3	75%
	MEK1	75%
	PTEN	80%
HepG2 (human liver)	MAPK1	80%
NIH 3T3-L1	MAPK1	70%
	MAPK3	70%
Secondary Human Astrocytes	Lamin A/C	80%
Primary Mouse Hepatocytes	ABC A1	70%
	Lamin A/C	81%

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.

Back to Top

 

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).

Back to Top