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ON-TARGETplus siRNA reagents reduce off-targets utilizing a patented dual-strand modification, while still providing guaranteed gene silencing of human, mouse and rat gene targets. This unmatched potency and specificity make ON-TARGETplus siRNA the premium choice for optimal knockdown and reduced off-targets. Simply search for your gene of interest and choose from the available product formats and quantities.
Our scientists and collaborators demonstrated in 2006† that siRNA off-targets are primarily mediated by antisense seed-region interactions, initiating the development of a dual-strand modification pattern to effectively reduce off-targets:
†Jackson et al., Position-specific chemical modification increases specificity of siRNA-mediated gene silencing. RNA12(7), 1197-1205 (2006).
siGENOME and ON-TARGETplus siRNA reagents (SMARTpool and three of four individual siRNAs) are guaranteed to silence target gene expression by at least 75% at the mRNA level when used under optimal delivery conditions (confirmed using validated positive control and measured at the mRNA level 24 to 48 hours after transfection using 100nM siRNA).
Note: Most siGENOME and ON-TARGETplus siRNA products are highly functional at 5 to 25nM working concentration.
Thermo Scientific offers three complete pre-designed product lines across human, mouse and rat genomes. Use the table below to assist you in determining the right siRNA product line for your needs.
Click the data figures below to learn more about ON-TARGETplus siRNA reagents.
ON-TARGETplus modifications reduce the overall number of off-targets and pooling reduces them even further
Only the ON-TARGETplus modification pattern addresses both siRNA strands for premium silencing
ON-TARGETplus siRNA dual-strand modification pattern reduces off-targets
Panels (A) and (B) are representative examples of off-target signatures with and without application of ON-TARGETplus modifications to (A) a single siRNA and (B) a SMARTpool reagent. Green bars indicate genes with 2-fold or more reduction of expression when treated with the indicated siRNA reagent.The ON-TARGETplus modifications reduced the off-targets when compared to unmodified siRNA. Pooling of siRNA and the ON-TARGETplus modification pattern independently, and in combination, provide significant reduction in off-target gene silencing. Panel (C) represents quantitation of off-targets (down-regulated by 2-fold or more) induced by the indicated siRNA reagents targeting 10 different genes (4 siRNAs per gene or a single SMARTpool reagent). Off-targets were quantified using microarray analysis (Agilent) then compiled. Each shaded box represents the middle 50% of the data set. Horizontal line in box: Median value of the data set. Vertical bars: minimum and maximum data values.
The ON-TARGETplus dual-strand chemical modification begins with the sense (passenger) strand being blocked from RISC uptake to favor antisense (guide) strand loading and reduce passenger strand-induced off-targets. However, the majority of siRNA off-targets are driven by the seed region of the guide strand. ON-TARGETplus is modified within its seed region to destabilize miRNA-like activity and improve specificity to the desired target for potent knockdown.
A 2006 publication demonstrates that off-target effects are primarily driven by antisense strand seed activity†. Therefore, sense strand inactivation alone does not decrease the total number of off-target genes.
ON-TARGETplus modifications account for both strands:
The ON-TARGETplus modification pattern dramatically reduces off-targets. Off-target effects induced by the indicated siRNAs were quantified using microarray analysis. For each target, three different siRNAs were used: unmodified, sense strand-inactivated, and ON-TARGETplus-modified. Data shown represents genes down-regulated by two-fold or more. HEK293 cells were transfected with 100 nM siRNA using 0.2 μL of DharmaFECT 1. Data was analyzed at 24 hours.