Human ON-TARGETplus siRNA Library

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Dharmacon Screening libraries
Human ON-TARGETplus siRNA Library - Genome

Human ON-TARGETplus siRNA Library - Whole 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 (*PP-0200 Echo Qualified 384-Well Polypropylene Microplate; 384PP).

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

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

(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.