BLOCK-iT™ Dicer Kits provide highly effective siRNA pools for RNAi knockdown. Diced siRNA (d-siRNA) pools are a cost-effective approach for completing multiple experiments with the same target. Experiments with Dicer provide robust knockdown and are particularly helpful in assessing the RNAi effects of several genes of interest or to follow up on microarray analysis. Once an RNAi effect is identified, additional evaluation can include synthetic or vector approaches.
Dicer technology is applicable for a variety of research needs
- Inhibiting genes recalcitrant to knockdown by other methods
- Studying multiple genes while avoiding the cost of ordering multiple synthetic sequences
- Running multiple transfection experiments in parallel by using the large amount of d-siRNA generated from each reaction
- Knocking down a target mRNA when identifying a specific target site has been difficult
- Generating a pool of d-siRNA that is unique to a gene or gene family to maximize the potential for exceptionally potent knockdown
Strong knockdown with d-siRNA pools generated by in vitro Dicer
The BLOCK-iT™ Complete Dicer RNAi Kit supplies all the technology and reagents needed to generate long dsRNA and cleave it into pools of d-siRNA. The kit includes everything required to generate, and purify d-siRNA pools, and transfect mammalian cells for strong gene inhibition (Figure 1).
Figure 1. Diced pools result in successful RNAi. Diced pools of siRNA (d-siRNA) were generated from β-galactosidase (β-gal) or luciferase dsRNA transcripts and used to determine the capability of dsiRNA to exclusively knock down gene activity. GripTite™ 293 MSR cells were cotransfected as follows: positive control plasmids pcDNA™1.2/V5-GW/lacZ and pcDNA™5/FRT/luc, expressing β-gal and luciferase reporters, respectively; positive control plasmids plus 50 ng of lacZ d-siRNA; or positive control plasmids plus luc d-siRNA. For each reporter, the corresponding d-siRNA successfully reduced the expression of reporter activity, whereas d-siRNA derived from the other transcript did
not affect expression.
Straightforward preparation of a potent d-siRNA mix
The BLOCK-iT™ RNAi Dicer Kits are the only systems available that contain recombinant Dicer enzyme, a high-quality d-siRNA purification module, and the most popular transfection reagent for RNAi, Lipofectamine™ RNAiMAX. Using in vitro dicing technology to achieve robust targeted inhibition of gene expression is easily accomplished by generating a pool of d-siRNA in three simple steps (Figure 2). It is not necessary to design and order synthetic oligos. Each kit contains everything needed to generate, from a PCR fragment of a target gene, a highly effective pool of siRNA. Larger custom sizes of the exceptionally pure recombinant Dicer enzyme are also available.
Figure 2. Three simple steps to successful RNAi with Dicer pools.