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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:

Expand All Tabs
Catalog Number Amount of reagent
409-01 0.1 ml
409-10 1 ml
409-50 5 x 1 ml
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).

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).

INTERFERin-fig1 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).

INTERFERin-fig2 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).

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).

INTERFERin-fig4 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 INTERFERin protocol 2012 2

Take a look at our Expert Tips in Genetic Engineering and Biotechnology News.

 

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.