Technical Note siRNA Transfection INTERFERin |
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- Great silencing using as little as 1 nM siRNA
- Over 90% gene silencing in a wide variety of cells
- Easy standard protocol
- Gentle mode of action for more robust data and excellent cell viability
- Compatible with serum and antibiotics
INTERFERin® provides more than 90% silencing efficiency at 1 nM siRNA in a wide variety of cells; hence avoiding off-target effects.
For High Throughput Screening Applications, Polyplus-transfection developed a dedicated reagent: INTERFERin®-HTS.
More information:
Ordering information
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1 ml of INTERFERin® is sufficient to perform 500-1000 transfections in 24-well plates. Bulk sizes available, please contact us. |
Several publications show that using low siRNA concentrations avoids off-target effects observed when transfecting siRNA at higher concentrations1,2. Ensure more reliable data by minimizing these unwanted non specific side-effects by using as little siRNA as possible. This is why INTERFERin® has been specifically designed to provide high silencing efficiency using low siRNA amount.
INTERFERin®-mediated delivery of 1 nM of a specific siRNA shows selective and highly efficient knockdown of gene expression, while a competitor (L2K) needs at least 10 nM siRNA to reach 50% silencing efficiency (Fig 1).
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Fig 1. INTERFERin® only requires 1 nM siRNA for efficient gene silencing. 3LL cells stably expressing firefly luciferase were transfected with an anti-Luc siRNA using INTERFERin® or competitor L2K according to the manufacturer’s recommendation. Luciferase expression was measured after 48 h using a conventional assay. No inhibition was observed with control siRNA. |
Amazingly, 50% silencing is still achieved at 10 pM siRNA (Fig. 2).
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Fig 2. INTERFERin® is highly efficient even at extremely low siRNA concentration. A549-GL3Luc cells were transfected in the presence of serum with decreasing concentrations of an anti-Luc siRNA using INTERFERin®. Luciferase expression was measured after 48 h using a conventional assay. No inhibition was observed with control siRNA. |
Transfection of 1 nM siRNA targeting endogenous lamin A/C with INTERFERin® drastically reduces lamin gene expression to barely detectable level (Fig. 3).
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Fig 3. Transfection of 1 nM siRNA with INTERFERin® results in efficient and specific gene silencing. CaSki cells were transfected with 1 nM lamin A/C siRNA using INTERFERin®. After 48 h, lamin A/C silencing efficiency was determined by immunofluorescence microscopy. |
For adherent cell lines or primary cells, 1 nM siRNA is sufficient to obtain more than 90 % gene silencing. For suspension cell lines, 80% silencing can still be reached by INTERFERin® using 5 nM siRNA (Table 1).
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Table 1. Successfully transfected cell lines and silencing efficiencies obtained with INTERFERin®. |
When it comes to cell viability, INTERFERin® outperforms other transfection reagents. 48 h after transfection with 1 nM siRNA, cells transfected with INTERFERin® appear healthy, while toxicity is clearly observed with reagent S (Fig. 4).
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Fig 4. INTERFERin® is extremely gentle on cells. Comparison of cell morphology 48 h after siRNA transfection using INTERFERin® or competitor reagents. A549-GL3Luc cells were transfected in the presence of serum with 1 nM Luciferase siRNA using INTERFERin® or competitors S or H according to the manufacturer’s protocol |
INTERFERin® is ready to use and the protocol is straightforward. Start with 1 nM siRNA as a starting concentration for silencing of most genes in most cell types (Fig. 5).
INTERFERin® is compatible with both serum and antibiotics, hence forget time consuming medium changes and washes; INTERFERin® can be left on the cells without any adverse effects.
| Fig 5. INTERFERin® standard protocolfor 24-well plate |  |
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1. Birmingham, A. et al., (2006) Nature Methods, Vol.3, No3, 199-204
2. Caffrey, D. et al., (2011) PLoS One. 2011;6(7):e21503. Epub 2011 Jul 5.
