Decode™ RNAi viral screening libraries combine the advantages of shRNAmir design, the power of viral delivery and the convenience of ready-to-use pooled viral particles to facilitate multiplexed RNAi screening. The high titer viral pool format enables RNAi screening without the cost and labor associated with individually arrayed screens. Perform genome-scale RNAi screens for genes regulating cellular responses, signaling pathways or discover genes with novel functions - all at the convenience of your benchtop.

Decode RNAi viral screening pool format is now available for the human GIPZ lentiviral shRNAmir library. Pooled screening libraries are available for the whole genome (7 pools of 10K constructs each) or only for highly characterized, annotated genes (3 pools of 10K constructs each).

Advantages of GIPZ lentiviral shRNAmir include:

• Efficient single copy knockdown
• Track shRNAmir expression with TurboGFP
  
• Lentiviral delivery extends RNAi screening to primary and non-dividing cells 
• Positive and negative selection screening kits available
  

Technical details

Positive and negative selection screening using Decode RNAi viral libraries

NEW! Negative selection screening kits include:

• Viral pools each containing 10K GIPZ lentiviral shRNAmir
• Non-silencing control shRNAmir construct-Lentiviral particles
• PCR primers for barcode amplification
• Positive control plasmid DNA for PCR amplification
• Two barcode microarrays for deconvolution of hits

Decode RNAi Positive selection screening kits include:

• Viral pools each containing 10K GIPZ lentiviral shRNAmir
• Non-silencing control shRNAmir construct-Lentiviral particles
• PCR primers for shRNAmir amplification
• Positive control plasmid DNA for PCR amplification
• Sequencing primer to identify shRNAmir

Decode RNAi viral screening pool specifications

Each Decode RNAi viral screening pool contains ~10,000 shRNAmir as viral particles. Viral titers of each pool are greater than 5x10^8TU/ml. Depending on the target cell line used one Decode RNAi viral screening kit contains enough virus to perform two screens in triplicate.

Decode RNAi Barcode microarray specifications

Each Decode RNAi barcode microarray consists of 2x105K arrays spotted with sequences unique to each shRNAmir using Agilent’s Sure Print technology (in situ base by base addition to slide). Barcode hybridizing probes have been optimized using Agilent algorithms for assessing probe quality (i.e. base composition, melting temperature etc.). Each array contains  58,498 probe sequences (most duplicated on the array) also includes Agilent controls. Details

Published multiplexed RNAi screens performed using Open Biosystems' shRNAmir libraries:

Positive selection RNAi screens

Westbrook T et al 2005 "A Genetic Screen for Candidate Tumor Suppressors Identifies REST" Cell, Vol. 121, 837–848.

Gazin C et al 2007 "An elaborate pathway required for Ras-mediated epigenetic silencing" Nature Vol 449: 1073-1077.

Gobeil S, et al 2008 "A genome-wide shRNA screen identifies GAS1 as a novel melanoma metastasis supressor gene" Genes and Development 22: 2932-2940.

Negative selection (barcode) RNAi screens

Silva JM et al 2008 " Profiling Essential Genes in Human Mammary Cells by Multiplex RNAi Screening" Science Vol 319, 617-620

Schlabach M et al 2008 "Cancer Proliferation Gene Discovery Through Functional Genomics" Science Vol 319, 620-624

This product is covered by the following limited use licenses:

Limited Use License-shRNA technologies
Limited Use License-ddRNAi
Limited Use License-TurboGFP