ON-TARGETplus GAPD Control Pool is an ideal positive control for RNAi experiments using SMARTpool reagents. ON-TARGETplus GAPD Control Pool is a pool of four highly functional, validated siRNAs for guaranteed silencing of GAPD in human, mouse, or rat cells. The ON-TARGETplus siRNA patented dual-strand modification pattern reduces off-targets by up to 90%, allowing improved specificity.
Also known as glyceraldehyde-3-phosphate dehydrogenase or GAPDH, GAPD is an important enzyme in carbohydrate metabolism that is well conserved across the animal kingdom. This gene is abundantly expressed in most cells and because it is non-essential, knockdown of the corresponding mRNA does not affect cell viability
- Ideal positive controls for RNAi experiments using human, mouse or rat SMARTpool reagents
- Pool of four highly functional, validated siRNAs for guaranteed silencing of GAPD in human, mouse, or rat cells
- Targets accession number NM_002046 (human) or NM_008084 (mouse) or XM_575242 (rat)
Only the ON-TARGETplus modification pattern addresses both siRNA strands for premium specificity
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 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.