Figure . Silencing potency of siRNAs containing ON-TARGET™ modifications. (A) Three different siRNAs, targeting human
cyclophilin B mRNA, were individually tested in a) unmodified, b) antisense strand modi-fied, and c) sense strand modified
forms for their ability to direct mRNA degradation in HEK293 cells. In all three cases, unmodified duplexes efficiently silenced the target. When ON-TARGET™ Modifications were applied to the antisense strand (S-*AS), duplex functionality was
eliminated, probably because the antisense strand (the strand responsible for directing gene-specific silencing) is unable to associate with the RISC. In contrast, when the same modifications are applied to the sense strand only (*S-AS), the silencing potency of the duplex is maintained or increased (ns = nonspecific; Control = non-targeting siRNA).(B) A heat map resulting from microarray studies illustrates the off-target expression profile of cells treated with unmodified and modified siRNAs directed against Gene A. Note that in this case, modification of the sense strand eliminates the majority of the off-target signature (illustrated primarily in the boxed region). When only the antisense strand is modified, off-target effects occur (blue = down-regulation, black = no effect, pink = up-regulation). (Heat map data presented with permission from A. Jackson and P.S. Linsley, Rosetta Inpharmatics LLC.) B.
The patented ON-TARGETplus modification pattern for specificity, combined with the SMARTselection algorithm for efficient, guaranteed target gene silencing, makes Thermo Scientific ON-TARGETplus siRNA the premium choice for optimal knockdown and reduced off-targets.
ON-TARGETplus SMARTpool and individual siRNAs are available as pre-designed, genome-wide reagents for human, mouse and rat. Simply search for your gene of interest and choose from the available product formats and quantities.
Highlights
• Off-targets reduced by up to 90% compared to unmodified siRNA
• Guaranteed silencing by SMARTpool and 3 of 4 individual siRNAs (see Specifications)
• Sequence information provided with siRNA purchase
A two-stranded mechanism requires a two-stranded solution
Dharmacon scientists and collaborators demonstrated in 2006 (see Reference 1) that siRNA off-targets are mediated by antisense seed-region interactions, initiating the development of a dual-strand modification pattern:
• Sense strand is modified to prevent interaction with RISC and favor antisense strand uptake
• Antisense strand seed region is modified to destabilize off-target activity and enhance target specificity
Seed-region analysis on siRNA designs reduces miRNA-induced off-targets
A landmark publication (see Reference 2) from the Thermo Scientific Dharmacon research group was the first to experimentally demonstrate the key role of the seed region in mediating off-targets. A subsequent 2008 paper (see Reference 3) showed the importance of seed frequency in the 3'UTR as in indicator of its likelihood to cause off-targets. These principles were subsequently applied to our siRNA designs to improve specificity:
• Design filters eliminate common seed regions likely to cause miRNA-like off-targets
• Seed frequency analysis for siRNA designs minimize off-target effects
Available Product Formats
•SMARTpool: A mixture of 4 siRNA provided as a single reagent; providing advantages in both potency and specificity. siGENOME and ON-TARGETplus guaranteed to silence by 75% or better (seeSpecifications).
•Set of 4: A convenient option for purchasing aliquots of all 4 individual siRNAs targeting a single gene. Three of four siGENOME and ON-TARGETplus siRNAs are guaranteed to silence by 75% or better (seeSpecifications).
•Set of 4 Upgrade: Discounted price on the Set of 4 based on a concurrent or prior purchase of the SMARTpool reagent for the same gene.
•Individual siRNAs: Select 1, 2, or 3 individual siRNAs per gene. Minimum purchase of 4 at the 2 nmol size.