TransIT-TKO® Transfection Reagent
A high efficiency, low toxicity, siRNA transfection reagent for mammalian cells
- Broad Spectrum siRNA 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.
Cell Types Successfully Transfected by Mirus Bio:
A549, BHK-21, BNL.CL2, C2C12, C6, CHO-KI, COS-7, Daoy, DB-TRG-05MG, D1-TNC1, DU145, HEK 293, HeLa, Hepa1c1c7, HepG2, human astrocytes, Jurkat, Keratinocytes (NIKS), MCF-7, Neuro-2a, NIH 3T3, PC-3, primary mouse hepatocytes, RAW 264.7, SK-N-MC, THP-1, Vero.
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. |
|
 |
| |
| 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. |
|
| |
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. |
|
 |
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). |
|
© 2008 Mirus Bio Corporation. All Rights Reserved.