Human ON-TARGETplus siRNA Library

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Dharmacon Screening libraries
Human ON-TARGETplus siRNA Library - Druggable Genome

Human ON-TARGETplus siRNA Library - Druggable Genome

siRNA designed and modified for greater specificity

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

The ON-TARGETplus Druggable Genome library contains the genes that are considered potential targets for therapeutics, including kinases, proteases, phosphatases, GPCRs, ion channels, ubiquitin ligases, and genes involved in senescence, autophagy, DNA repair, and characterized nuclear receptors, This extensive library is delivered as nine distinct subset siRNA libraries in the 96-well format. In the 384-well format the first eight subsets are plated continuously to reduce partial plates.

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- or 384-well plates
Guaranteed target gene knockdown (see Specifications tab)

Nine subsets make up the human druggable genome siRNA Library

GPCR
Protein Kinases
Ion Channels
Phosphatases
Drug Targets- Includes genes involved with apoptosis, senesence, nucleic acid binding, autophagy, DNA repair, and characterized nuclear receptors.

Proteases
Ubiquitin Conjugation 1 - Cullins, E1, E2, HECT E3 Ligases
Ubiquitin Conjugation 2 - F-box, SOCS box E3 Ligases
Ubiquitin Conjugation 3 - RING finger and RING finger-like E3 Ligases

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 upon request (*PP-0200 Echo Qualified 384-Well Polypropylene Microplate (384PP)).

Druggable Genome siRNA Library

The Druggable Genome library contains the genes that are considered potential targets for therapeutics, and is delivered as nine distinct subset siRNA libraries in the 96-well format. In the 384-well format the first eight subsets are plated continuously to reduce partial plates. The Drug Targets portion includes genes involved with apoptosis, senesence, nucleic acid binding, autophagy, DNA repair, and characterized nuclear receptors.

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.