Small interfering RNA (siRNA) has recently emerged as a novel tool in the functional genomics arena of small RNA molecules (siRNA and microRNA). RNA interference is a mechanism of gene silencing at the mRNA level. This phenomenon is triggered by small interfering (si)RNAs and micro (mi)RNAs. These molecules involved in gene regulation belong to an expanding class of small non-coding RNAs. siRNAs are capable of inhibiting gene expression by either directing the degradation of homologous mRNA targets or inducing the repression of translation of mRNA targets.
In 2002, siRNA was hailed by Science magazine as being the "Breakthrough of the year" technology. In RNAi experiments, the most critical design factor is specific target recognition which is critical because the efficiency level of siRNA is different for each site. Silencing the correct gene enables researchers to obtain reproducible experimental results which can lead to the subsequent use of siRNA as a genetic drug. Experimental success depends upon several factors. The most critical among these factors is the design of effective and specific siRNA. Bioneer, in collaboration with the world renowned National Genome Information Center (NGIC) at KRIBB institute, has developed a proprietary siRNA selection algorithm. Turbo si-Designer identifies highly effective siRNA target sites with exceptional success rates. Several important parameters including base composition, the number of repetitive bases in a row, thermodynamic instability, energy profiling and base preference were considered in the development of Turbo si-Designer. The siRNAs spanning SNP sites are removed and non-specific siRNAs are eliminated after BLAST to minimize off-target effects. The resulting candidates are then ranked according to the NGIC scoring system. The performance of the algorithm was evaluated by designing hundreds of siRNAs and testing the siRNA nockdown efficacy by Real-time PCR analysis. Over 80% of the siRNAs tested showed > 75% knockdown of the target mRNA and more than 40% of siRNAs induced > 90% knockdown. Notably, siRNAs with the low NGIC score were mostly nonfunctional, indicating that ineffective siRNAs are efficiently removed by Turbo si-Designer.
To validate the performance efficacy of Turbo si-Designer, Bioneer tested the knockdown efficiency of 82 predesigned siRNAs in anti-apoptosis and cell division related genes (Survivin). The siRNA was transfected into A549 lung carcinoma cells and the knockdown efficiency was then analyzed using QuantiGene ViewRNA Analysis. As seen on the Figure 1B., the lower-scoring siRNAs are not effective compared to the higher-scoring siRNA (Figure 1A.) and thus Turbo si-Designer can predict the higher efficiency siRNA by the exclusion of ineffective siRNA sites (see Figure 1).

Figure 1. siRNA Knockdown efficiency of siRNAs designed by Turbo si-Designer was analyzed by Northern blot and real-time PCR analysis.
A) siRNA Knockdown efficiency of high NGIC score siRNAs.
B) siRNA Knockdown efficiency of low NGIC score siRNAs.

Since all siRNAs are not equally potent and not all silencing is gene specific, Bioneer strongly suggests performing siRNA Knockdown efficiency experiments with at least 3 siRNAs per target gene. Bioneer guarantees 80% siRNA Knockdown efficiency when purchasing 3 siRNAs for the same target gene.

Bioneer 80% Guarantee Policy

When a customer purchases 3 siRNAs for one target gene, Bioneer guarantees 80% reduction in the target mRNA level for at least 2 siRNAs. If less than 80% reduction is shown for more than 2 siRNAs, Bioneer will design and provide 2 new siRNAs free of charge.*
*Bioneer reserves the right to request the customer his/her supporting data inclusive of:
1. Transfection efficiency data: With NC (AccuTarget™ Fluorescein-labeled Negative Control) and siRNA concentration at 100 nM each
2. siRNA knockdown efficiency data: PC (AccuTarget™ GAPDH/GFP/Luciferase siRNA) and NC (AccuTarget™ Negative Control) at 100 nM each

Figure 2. siRNA Knockdown efficiency of AccuTarget Genome-wide Predesigned siRNA. AccuTarget Predesigned siRNAs are highly effective. To determine siRNA Knockdown efficiency of predesigned siRNAs, HeLa cells were transfected with siRNAs at 100 nM concentration. 24 hours post-transfection, total RNA was isolated and the level of target mRNA was measured by qRT-PCR. This data demonstrates the effectiveness of the Turbo si-Designer algorithm: 83.8% of tested siRNAs induced >70% siRNA Knockdown and 38.1% of tested siRNAs elicited >90% knockdown.

Notice to Purchaser

All siRNA Products: For Research Use Only. Not For Use in Diagnostic Procedures.

Limited License

This product is licensed under European Patents 1144623, 121945 and foreign equivalents from Alnylam Pharmaceuticals, Inc., Cambridge, USA and is provided only for use in academic and commercial research whose purpose is to elucidate gene function, including research to validate potential gene products and pathways for drug discovery and development and to screen non-siRNA based compounds (but excluding the evaluation or characterization of this product as the potential basis for a siRNA-based drug) and not for any other commercial purposes. Information about licenses for commercial use (including discovery and development of siRNA-based drugs) is available from Alnylam Pharmaceuticals, Inc., 300 Third Street, Cambridge, MA 02142, USA.

This product is sold for research use only and is not to be administered to humans or used for medical diagnostics. Buyer acknowledges and agrees that all intellectual property rights in the products (including, without limitation, the siRNA sequences used to create such products) and in any Bioneer technology, intellectual property and know-how used to make or useful for the manufacture or use of the products will at all times remain vested in Bioneer (other than any ownership interest that buyer may have in non-public proprietary target genes supplied by buyer to Bioneer in connection with custom products).

Trademark: AccuTarget is a trademark of Bioneer Corporation.