ON-TARGETplus siRNA

ON-TARGETplus siRNA

Product Overview

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ON-TARGETplus siRNA

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.

Highlights

  • We now offer a new 2 nmol size when purchasing pre-designed ON-TARGETplus or siGENOME siRNAs for human, mouse, or rat.
  • Off-targets reduced by up to 90% compared to unmodified siRNA.
  • Guaranteed silencing by SMARTpool and 3 of 4 individual siRNAs (See Product 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).

Product Formats

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.

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, 3 or 4 individual siRNAs per gene.

†Our siRNA knockdown guarantee

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.

Ordering Guidelines

Approximate # reactions (wells) at 25 nM siRNA concentration (assuming no loss from pipetting)

nmol		 96-well plate
(100 uL total reaction volume)		 24-well plate
(500 uL total reaction volume)		 12-well plate
(1000 uL total reaction volume)
2 800 160 80
5 2000 400 200
10 4000 800 400
20 8000 1600 800

Product Line Selection

Which siRNA is right for you?

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.

Cost-effective, efficient silencing High specificity for reduced off-targets + efficient silencing Target silencing in difficult-to-transfect cells
siGENOME siRNA ON-TARGETplus siRNA Accell siRNA
Pre-designed for Human, Mouse and Rat protein-coding genes + + +
Recommended for transfectable mammalian cells in culture + +
Recommended for neuronal, suspension, primary and other difficult-to-transfect cells +
Recommended transfection reagent DharmaFECT transfection reagent DharmaFECT transfection reagent None required
Available as SMARTpool reagent + + +
Available as four individual siRNAs + + +
New 2 nmol size available for individual siRNA + +
Guaranteed knockdown by SMARTpool and 3 of 4 siRNAs + +
Sense strand is modified to prevent interaction with RISC and favor antisense strand uptake Selective application when thermodynamic analysis indicates it necessary for favorable antisense RISC loading + +
Antisense strand seed region is modified to destabilize off-target activity and enhance target specificity + +
Modifications to facilitate cellular uptake without separate transfection reagents +
Stabilizing modifications to prevent nuclease-mediated degradation +
Sequence information provided with purchase + + +

Supporting Data

Click the data figures below to learn more about ON-TARGETplus siRNA reagents.

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on-target plus heatmap boxplot

ON-TARGETplus modifications reduce the overall number of off-targets and pooling reduces them even further

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on-target plus modification pattern

Only the ON-TARGETplus modification pattern addresses both siRNA strands for premium silencing

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on-target plus reduces off-target effects

ON-TARGETplus siRNA dual-strand modification pattern reduces off-targets

ON-TARGETplus modifications reduce the overall number of off-targets and pooling reduces them even further

heatmap boxplot otp

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

otp modification pattern

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

otp reduces off-target effects

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.