ON-TARGETplus siRNA Reagents - Mouse

siRNA designed and modified for greater specificity
Now part of GE Healthcare - Learn More


Guaranteed gene silencing and patented modifications to reduce off-targets; available as pre-designed ON-TARGETplus SMARTpool and siRNA reagents for human, mouse or rat.
Enter a common gene identifier and click SEARCH to find products for your gene

Common gene identifiers for your gene can be found at NCBI Gene or miRBase.

IdentifierDescriptionExamples
Gene IDThe Gene ID is a unique identifying number for each gene in the RefSeq database and tends to be the most stable gene identifier.
  • 672
  • 22421
Accession NumberAccession Numbers are identifiers for a sequence record. The preferred accession numbers for RNAi products are found in the Reference Sequence (RefSeq) database, a non-redundant, curated, and annotated collection of sequence records for major model organisms. RefSeq accession numbers for mRNA begin with the characters NM_ or XM_ followed by six or nine digits. For gene expression products, the GenBank accession number for the clone can also be used for more specific search results.
  • NM_001798
  • NM_001170537
  • BC068303
Gene SymbolThe Gene Symbol can be the official or alternate symbol as displayed on NCBI Gene.
  • BRCA1
  • CDK1
  • CTNNB1
Gene DescriptionThe Gene Description is also known as the Official Full Name on NCBI Gene.
  • myogenic differentiation 1
  • thyroid peroxidase
  • breast cancer 1, early onset
miRBase IdentifiersmiRBase accession numbers are the most stable identifiers for microRNA. The miRBase ID, or name, may also be used.
  • miRBase ID: hsa-let-7d, mmu-mir-122
  • Precurser Accession: MI0000065, MI0000256
  • Mature Accession: MIMAT0000065, MIMAT0000246

If you require further assistance with your search, please contact Technical Support at ts.dharmacon@ge.com

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ProductCatalog Number 
ON-TARGETplus SMARTpool - Mouse
L-MOUSE-XX-0005 5 nmol - $445.00select
ON-TARGETplus Set of 4 Upgrade - Mouse
LU-MOUSE-XX-0002 2 nmol - $425.00select
ON-TARGETplus Set of 4 - Mouse
LQ-MOUSE-XX-0002 2 nmol - $730.00select
ON-TARGETplus siRNA - Mouse
J-MOUSE-XX-0002 2 nmol - $182.00select

The patented ON-TARGETplus modification pattern for specificity, combined with the SMARTselection algorithm for efficient, guaranteed target gene silencing, makes 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 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.
  
HazardousNo
Shelf Life12 Months
Shipping ConditionAmbient
Storage Condition-20 C

Order Quantity Guidelines

ON-TARGETplus reagents are routinely used at 5 to 25 nM concentration. The calculations below, based on 25 nM, are for estimation purposes only and assume no loss from pipetting.

  Approximate # reactions (wells) at 25 nM siRNA concentration
nmol 96-well plate
(100 µL total reaction volume)
24-well plate
(500 µL total reaction volume)
12-well plate
(1000 µL total reaction volume)
1 400 80 40
2 800 160 80
5 2000 400 200
10 4000 800 400
20 8000 1600 800

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 100 nM siRNA).

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

False phenotypes due to off-targets are alleviated by ON-TARGETplus SMARTpool reagents while target gene knockdown is maintained

False phenotypes due to off-targets are alleviated by ON-TARGETplus SMARTpool reagents while target gene knockdown is maintained

False phenotypes due to off-targets are alleviated by ON-TARGETplus SMARTpool reagents while target gene knockdown is maintained

The effect of silencing ARPC1B on cell migration was studied in a breast cancer cell line. A monolayer of cells was uniformly scraped and the rate of cell migration to close the scrape (wound healing) was evaluated. Both unmodified and ON-TARGETplus siRNA reagents induced potent target knockdown. Inconsistent phenotypes due to off-target effects (red outline), were observed for cells transfected with unmodified individual siRNAs. The unmodified SMARTpool improved the false phenotype considerably, while the ON-TARGETplus SMARTpool significantly reduced off-target effects to produce a consistent phenotype.

In collaboration with Kaylene Simpson, Laura Selfors and Joan Brugge, Harvard Medical School.


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

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

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

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

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.


siRNA designs with low-frequency seed regions ensure fewer off-targets

siRNA designs with low-frequency seed regions ensure fewer off-targets

siRNA designs with low-frequency seed regions ensure fewer off-targets

ON-TARGETplus siRNA designs leverage sophisticated bioinformatics to reduce the likelihood of miRNA-like off-targets from high-frequency or highly conserved miRNA seed regions. siRNAs with low seed frequency have a significantly lower number of off-targets than siRNAs with medium or high frequency seeds. Five siRNAs with low, medium, or high frequency seed regions were transfected into HeLa cells and their associated off-target signatures assessed via global expression profiling (Agilent 22K platform). siRNA sequences were constant at positions 1 and 8-19, only the seed regions (positions 2-7) were altered.

Low frequency seeds: < 350 occurrences in the HeLa transcriptome

Medium frequency: 2500-2800 occurrences

High frequency: >3800 occurrences


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

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

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.


References

  1. A. L. Jackson et al., Position-specific chemical modification increases specificity of siRNA-mediated gene silencing. RNA. 12(7), 1197-1205 (July 2006).
  2. A. Birmingham et al., 3'-UTR seed matches, but not overall identity, are associated with RNAi off-targets. Nature Methods. 3(3), 199-204 (March 2006).
  3. E. M. Anderson et al., Experimental validation of the importance of seed complement frequency to siRNA specificity. RNA. 14(5), 853-861 (May 2008).