ON TARGETplus siRNA

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 

• Off-targets reduced by up to 90% compared to unmodified siRNA.
• Guaranteed silencing by SMARTpool and 3 of 4 individual siRNAs (see formats tab).
• Sequence information provided with siRNA purchase.

A two-stranded mechanism requires a two-stranded solution

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:

• 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.

†Jackson et al., Position-specific chemical modification increases specificity of siRNA-mediated gene silencing. RNA12(7), 1197-1205 (2006).

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.

ON-TARGETplus positive control reagents

• ON-TARGETplus cyclophilin B control siRNA

  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.

• ON-TARGETplus cyclophilin B control pool

  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.

• ON-TARGETplus GAPD control siRNA

  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.

• ON-TARGETplus GAPD control pool

  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.

ON-TARGETplus negative control reagents

• ON-TARGETplus non-targeting siRNAs

  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.

• ON-TARGETplus non-targeting pool

  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.

ON-TARGETplus siRNA libraries

• Human siRNA libraries

  Find the predefined gene family, including Druggable genes or Whole Genome, that’s right for your discovery efforts.

• Mouse siRNA libraries

  A wide selection of predefined siRNA libraries, including the most up-to-date Whole Mouse genome collection available.

• Cherry-pick libraries

  Fast and easy online configuration and ordering of plated siRNA & microRNA reagents targeting your genes of interest.

Custom siRNA synthesis

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.

SMARTpool:

• A mixture of 4 siRNA provided as a single reagent; providing advantages in both potency and specificity. siGENOME and ON-TARGETplus are guaranteed to silence by 75% or better.

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. 

Individual siRNAs:

• Select 1, 2, 3 or 4 individual siRNAs per gene.

Our siRNA knockdown guarantee

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 demonstrated to have been 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).

Ordering guidelines

Note: Most siGENOME and ON-TARGETplus siRNA products are highly functional at 5 to 25nM working concentration.

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:

• Sense strand is modified to prevent interaction with RISC and favor antisense strand uptake
• Antisense strand seed region is modified to minimize seed-related off-targeting

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.