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Products > RNAi (RNA Interference) > Pooled shRNA Screening Libraries > Product Overview

shERWOOD UltramiR Pooled Screening Libraries

Choice of promoter, reporter, gene content and more

Achieve superior RNAi screens with enhanced potency, sequence-verified  shRNA pools and flexible delivery strategies. 

Greater confidence in your results
  • Every clone is sequence verified – eliminate unwanted background from mutations introduced in chip-based pools
  • More potent shRNA per gene – decrease false positive and false negative results
  • Equimolar pooling process - reduces variation between samples
Choose the best promoter for your cells
 
  • Select the optimal promoter for your target cell line with a choice of promoters- Choose from SFFV, hCMV, mCMV, hEF1-alpha, mEF1-alpha and more
Your choice of format
  • In vitro or in vivo pool formats.  Pools can be subdivided into minipools for lower complexity and higher representation for in vivo or other applications that require less complex pools
  • Choice of gene coverage - premade gene families or custom gene sets
  • Ready to use lentiviral particles or plasmid DNA format
Pool Deconvolution Service
 
  • NGS analysis service - send us your genomic DNA and receive counts for every shRNA construct

Promoter Selection Kit

Mammalian promoters may differ in expression level or be silenced over time depending on the target cell line.  Variation in expression level can affect fluorescent marker expression as well as knockdown efficiency.  For cell lines where the optimal promoter is unknown, the pZIP promoter testing kit includes ready-to-use lentiviral particles expressing ZsGreen or RFP from five different promoters (mCMV, hCMV, SFFV, mEF1-alpha, and hEF1-alpha). The fluorescent marker and shRNA are on the same transcript allowing a quick visual assessment of expression efficiency.

The promoter selection kit can be used to test shRNA expression in hard to transfect cells to select the promoter that produces optimal expression.Target gene sets or pooled shRNA libraries can be ordered in a choice of promoters, reporters and vectors.

Visualizing expression levels from different promoters
(hCMV, mCMV and SFFV) in D2.OR cells. Cells are transduced at
similar titers. Fluorescence reflects expression levels of transcript
which includes the shRNA-mir.

The pZIP promoter selection kit provides sufficient viral particles (25 ul of 10^8 TU/ml) of a non-targeting shRNA in each of five promoter/vector (mCMV, hCMV, SFFV, mEF1-alpha and hEF1-alpha) options to transduce your target cell line and determine the optimal promoter for shRNA expression. Simply use the provided pre-packaged viral particles of the panel of vectors expressing RFP or ZsGreen from the different promoters to easily detect expression efficiency.


Lentiviral shRNA Vector Design
Included in kit: 25 ul of 10^8 TU/ml of each murine CMV, human CMV, SFFV, hEF1-alpha and mEF1-alpha vectors with your choice of fluorescent marker - RFP or ZsGreen

Flexible pool formats for optimal pooled shRNA screens:

In vitro, in vivo and ex vivo screening strategies have different requirements for pool formatting.  In vivo and ex vivo screens require smaller, less complex pools to facilitate a higher shRNA representation while in vitro screens can be efficiently performed with higher complexity pools.  Pooled shRNA screening libraries are available as viral particles that are ready-to-screen or plasmid DNA that can be packaged as needed for small or large-scale screens. In addition, we can subdivide into mini pools of ~1000 shRNA per pool to enable low complexity and higher representation for in vivo and other screening applications. 

Custom Pooled shRNA Libraries
Custom pools can be created from our shERWOOD UltramiR shRNA libraries and we can deliver any bulk volume. We can even achieve titers up to 10^9 TU/ml when requested. Choose the number of shRNAs for your pool to satisfy your in vitro or in vivo screening, and choose the optimal promoter and fluorescent reporter.  Contact us at info@transomic.com for more information and pricing.

Screening capacity:
 

  • Lentiviral pooled shRNA libraries provided as viral particles include enough virus to perform a screen at 500-1000-fold representation with at least 2 biological replicates.
  • Libraries provided as plasmid DNA include enough plasmid for more than 3 packaging reactions.  Total viral particles generated will depend on packaging efficiency, but is expected to provide enough viral particles to perform a screen in multiple cell lines at 500-1000-fold representation with 3 biological replicates.
  • Mini pool formats available upon request.

NGS Deconvolution Service

Need help deconvoluting your pooled screen? Send us your genomic DNA samples for sequencing and analysis.

A cost effective way to simplify your pooled RNAi screen analysis.  Send us the genomic DNA from your screen and we will return counts for each shRNA.  Our optimized protocols provide reproducibility and a quick turnaround time (4-6 weeks).

• Simplify your screen - Sequencing service including library preparation, next generation sequencing and data analysis
• Cost effective - Multiplex sequencing allows analysis of multiple samples in parallel
• Quick turnaround - 4-6 weeks from receipt of samples to delivery of data
• More data reproducibility - Optimized protocols and reagents for shRNA specific sequencing

 

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OMNIfect Transfection Reagent

Need a gentle and efficient transfection reagent for plasmid transfection?

OMNIfect™ transfection reagent is an easy-to-use, cationic delivery reagent providing high efficiency transfection of plasmid DNA coupled with low cytotoxicity across a broad range of cells. OMNIfect has been tested across a variety of cell lines and has proven to successfully transfect both adherent and suspension cell lines. 
  • Efficient delivery – high transfection efficiency with or without serum
  • Low cytotoxicity gentle on sensitive cell lines
  • Broad range of cells tested – including neuronal, stem and suspension cells
  • Easy-to-use protocols