| Tag Line Label |
| Tag Line Sub |
| |
|
Advanced Modifications for siRNA
|
|
|
Available on Custom siRNA Synthesis
|
Available on Custom OR
Predesigned siGENOME Collection
|
| |
ON-TARGET®
|
siSTABLE®
|
ON-TARGETplus™
|
|
Sense strand inactivation for reduced off-target silencing
|
√
|
√
|
√
|
|
Enhanced stability in nuclease-rich environments
|
|
√
|
|
|
Improved specificity to target gene
|
|
|
√
|
|
APPLICATIONS
|
• Silencing of highly conserved sequences
• Follow-up screening to eliminate false positives |
• in vivo RNAi-based target identification and validation
• Experiments in nuclease-rich environments |
• Screening
• Pathway analysis
• Target identification
• Highly conserved target silencing |
|
|  |
| |
A recent publication demonstrates that off-target effects are primarily driven by antisense strand seed activity.1 Therefore, sense strand inactivation alone does not decrease the total number of off-target genes.
ON-TARGETplus siRNA has the only modification pattern that addresses off-target effects caused by both strands.
• Sense strand is modified to prevent interaction with RISC and favor antisense strand uptake
• Antisense strand seed region is modified to minimize seedrelated off-targeting
1Jackson, A.L. et al. "Position-specific Chemical Modification Increases Specificity of siRNA-mediated Gene Silencing." RNA 12.7 (2006) 1197-1205.
|
|
| |
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.
|
|
|
| Panel 1 | |
| The stability of conventional siRNAs is acceptable for most basic research and screening applications in vitro. Unfortunately, the relatively short bioavailability of conventional siRNA may limit the potential of siRNA-mediated silencing in vivo. Conventional siRNA is degraded within minutes in serum-containing environments, making in vivo use of siRNA problematic. |
The data in Figure 1 show that the siSTABLE modification pattern dramatically extends the half-life of siRNA in the presence of 100% human serum as compared to the stability-enhanced siRNA from other commercial sources.
Additional Highlights
• Includes sense strand inactivation (ON-TARGET® modification) - minimizes off-target silencing and enhances specificity.
• Enhanced stability with no effect on cell viability.
• Reduced stress response compared to unmodified siRNA.
Controls
siSTABLE Controls have been well-characterized and are recommended for use with siSTABLE modified siRNAs.
More information here |
|
 |
|
|
|
| Panel 3 | |
| |
To address false-positive results due to sense-strand uptake, Dharmacon has developed proprietary chemical modifications which:
| |
• Exploit biochemical/biophysical factors to disrupt the sense strand-RISC interaction
• Inhibit activity of the sense strand while the antisense strand remains fully functional
• Minimize off-target activity |
|
|
|
| |
| |
Off-Target Effects
The functionality of an siRNA is based on the interaction of its antisense strand with the target mRNA and the RISC complex.
It is important to recognize that the siRNA sense strand also has the potential to interact with the RISC and an unintended mRNA target. This unintended silencing is known as an off-target effect.
The ON-TARGET chemical modification inhibits sense-strand uptake by RISC, thus reducing the potential for sense strand-mediated off-target effects. |
 |
|
|
|