siRNA designed and modified for greater specificity. 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).
Begin a gene search to place an order for human, mouse or rat ON-TARGETplus siRNA.
Find the predefined gene family, including Druggable genes or Whole Genome, that’s right for your discovery efforts.
A wide selection of predefined siRNA libraries, including the most up-to-date Whole Mouse genome collection available.
Human and mouse SMARTpool siRNA libraries in a novel pre-dispensed format to enable RNAi screening without the need for high-throughput automation.
Fast and easy online configuration and ordering of plated siRNA & microRNA reagents targeting your genes of interest.
No pre-designed product to fit your needs? Use our online design tools and extensive synthesis options to create a custom siRNA specific for your application.
ON-TARGETplus Control siRNAs and pools are produced with the patented ON-TARGETplus modification pattern for optimal specificity. They are recommended as controls for any siRNA experiment due to their minimized off-target effects. Select a species-specific positive control to silence an expressed housekeeping gene in your experimental cell type. Our panel of Non-targeting controls permits assessment of potential non-specific effects, to find the optimal negative control for your cell type, and your assay. Pooled siRNA controls are recommended for further reduction of off-targets, and for use with SMARTpool siRNA reagents.
A validated positive control siRNA targeting the cyclophilin B (PPIB) gene in human, mouse, or rat. Includes patented ON-TARGETplus modifications to minimize off-target effects. Useful for determination of optimal RNAi conditions.
A validated positive control pool of four siRNAs targeting the cyclophilin B (PPIB) gene in human, mouse, or rat. Includes patented ON-TARGETplus modifications to minimize off-target effects. Useful for determination of optimal RNAi conditions.
A validated positive control siRNA targeting the GAPD gene in human, mouse, or rat. Includes patented ON-TARGETplus
modifications to minimize off-target effects. Useful for determination of optimal RNAi conditions.
Validated positive control pool of four siRNAs targeting GAPD gene in human, mouse, or rat. Includes patented ON-TARGETplus modifications to minimize off-target effects. Useful for determination of optimal RNAi conditions.
Negative control siRNAs designed and microarray tested for minimal targeting of human, mouse, or rat genes. ON-TARGETplus modifications reduce potential off-targets. Recommended for determination of baseline cellular responses in RNAi experiments.
Negative control pool of four siRNAs designed and microarray tested for minimal targeting of human, mouse or rat genes. ON-TARGETplus modifications reduce potential off-targets. Recommended for determination of baseline cellular responses in RNAi experiments.
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.
Dharmacon offers four 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.
ON-TARGETplus modifications reduce the overall number of off-targets and pooling reduces them even further
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.
Only the ON-TARGETplus modification pattern addresses both siRNA strands for premium silencing
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.
ON-TARGETplus siRNA dual-strand modification pattern reduces off-targets
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.