Human ON-TARGETplus siRNA Library - Genome

siRNA designed and modified for greater specificity


An arrayed collection of siRNA reagents for RNAi screening, targeting the whole human genome. ON-TARGETplus siRNA is guaranteed to silence, and is modified to reduce off-targets for fewer false positives.

Genome-wide siRNA screening has become an invaluable discovery tool, and siRNA products are more often the choice for high quality, reliable screening libraries. The Human ON-TARGETplus Human Genome siRNA Library targets all unique human genes (NM Accession numbers) in the NCBI RefSeq database.

The ON-TARGETplus siRNA designs and modifications reduce off-target effects while maintaining high silencing potency for high-confidence screening results.

Highlights

  • Patented dual-strand ON-TARGETplus modification pattern on all siRNAs to 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
  • Available as SMARTpool siRNA reagents or a Set of 4 siRNAs in 96-well plates
  • Guaranteed target gene knockdown (see Specifications tab)
To support high-throughput screening environments, pre-defined and custom siRNA libraries in 384-well format may be provided in plates compatible with the Labcyte Echo acoustic liquid handler (*P-05525  Echo Qualified 384-Well Polypropylene Microplate; 384PP).
  
HazardousNo
Shelf Life12 Months
Shipping ConditionAmbient
Storage Condition-20 C

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.

Whole Genome siRNA Library Organization

Whole Genome siRNA Library Organization

Whole Genome siRNA Library Organization

The ~18,000 genes of the ON-TARGETplus Human Genome siRNA Library are organized according to the indicated subsets.


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

(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.


References

  1. B.D. Parsons, A. Schindler, D.H. Evans, E. Foley, A direct phenotypic comparison of siRNA pools and multiple individual duplexes in a functional assay. PLoS One. 4(12), e8471 (2009).
  2. M. Jiang, R. Instrell, B. Saunders, H. Berven, M. Howell, Tales from an academic RNAi screening facility; FAQs. Brief Funct. Genomics. 10(4), 227-237 (2011). [doi: 10.1093/bfgp/elr016]
  3. J. Borawski, A. Lindeman, F. Buxton, M. Labow, L.A. Gaither, Optimization procedure for small interfering RNA transfection in a 384-well format. J. Biomol. Screen. 12(4), 546-559 (2007).

Citations

  1. T. Ratovitski, E. Chighladze, E. Waldron, R. R. Hirschhorn, C. A. Ross, Cysteine proteases bleomycin hydrolase and cathepsin Z mediate N-terminal proteolysis and toxicity of mutant huntingtin. J. Biol. Chem. 286(14), 12578-12589 (2011). [Human Proteases]