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shERWOOD-UltramiR shRNA

shERWOOD-UltramiR shRNA

shERWOOD-UltramiR shRNA Reagents

shERWOOD shRNA Design + Optimized shRNA Processing = Superior Knockdown

shERWOOD-UltramiR shRNA reagents are next generation vector-based RNAi triggers designed using the proprietary shERWOOD algorithm developed and validated in Dr. Gregory Hannon’s laboratory at Cold Spring Harbor Laboratory (Knott et al 2014). An alternate microRNA scaffold "UltramiR" has been optimized for increased shRNA processing and potency based on new information on the key determinants for primary microRNA processing (Auyeung et al 2013).

The shERWOOD algorithm is based on the functional testing of over 250,000 shRNA sequences using a high-throughput sensor assay (Knott et al 2014) and uses key sequence characteristics for predicting shRNA potency to select the rare shRNA designs that are potent at single copy representation in the genome. The shERWOOD algorithm designs have been applied to the creation of  shERWOOD-UltramiR shRNA libraries targeting human, mouse and rat genomes.

Learn more about the shERWOOD algorithm.
Highlights:
  • shERWOOD algorithm designs - superior knockdown performance relative to existing shRNA design
  • UltramiR scaffold - increases shRNA processing and potency
  • All transcripts for a gene are targeted for the most complete knockdown
  • 100% Guaranteed* - Every shRNA is guaranteed to knockdown its intended target
Use the FETCH my gene® search tool to find and purchase lentiviral or retroviral shRNA targeting your gene of interest.

*All shRNA constructs in a target gene set are guaranteed to knockdown mRNA expression by >70%. Cell line of choice should demonstrate expression of the target gene using the non-targeting controls and should demonstrate gene knockdown using positive control shRNA (targeting PTEN or GAPDH).

Vectors and Specifications


Lentiviral, inducible lentiviral and retroviral vector options with a choice of promoters for optimal expression in your target cell lines.

The shERWOOD-UltramiR shRNA libraries are offered in a choice of promoters and fluorescent marker for optimal expression, and can be delivered by transfection or transduction.  shERWOOD-UltramiR shRNA can be purchased as target gene sets, custom focus sets, pooled screening libraries and whole genome. 

  • Deliver to a wide range of cell types including primary and non-dividing
  • Fluorescent marker allow direct visual detection of shRNA expression
  • Inducible or constitutive promoter options
  • Easily create stable cell lines with efficient integration and antibiotic selection

Purchase OMNIfect transfection reagent designed for efficient and gentle plasmid delivery.
 

Vector Options:
Lentiviral shRNA Vector Design

pZIP Lentiviral Vectors
(Click image for detailed vector information)
The pZIP lentiviral vector is available with six different promoter options (murine CMV, human CMV, SFFV, hEF1-alpha, mEF1-alpha and TRE3G) driving expression of shRNA. The fluorescent marker and shRNA are on the same transcript allowing the level of fluorescence in the cell to be used as a direct indication of shRNA expression through visual inspection. (See below the promoter selection kit to find out more information about the promoter best suited for your experiment.)
Lentiviral Inducible shRNA Expression Vector

pZIP Inducible Lentiviral Vector
(Click image for detailed vector information)
 
Retroviral shRNA-mir Vector
 
pLMN Retroviral Vector
(Click image for detailed vector information)
 


pLMPd-Ametrine Retroviral Vector
(Click image for detailed vector information)
 

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

shERWOOD-UltramiR shRNA Target Gene Sets

shERWOOD-UltramiR shRNA target gene sets target human, mouse and rat genes. The target gene sets come with the murine CMV promoter expresses efficiently in a number of human and mouse cell lines. However, for cell lines where a different promoter may be optimal, the pZIP promoter selection kit is available to quickly test for optimal expression in your target cell line.

Target gene set includes:

  • 3 top ranked shRNA targeting a gene of interest
  • Non-targeting control
  • Available as bacterial glycerol stocks or viral particles
  • Inducible option available
Use FETCH my gene search tool to find shERWOOD-UltramiR shRNA targeting your gene of interest.

shERWOOD-UltramiR shRNA Custom Focused Sets

shERWOOD-UltramiR shRNA is available targeting predefined gene families, pathways or your custom gene list. The custom sets can be purchased as arrayed or pooled formats. Use the request a quote button below to inquire about pricing and available predefined focused sets or send your custom gene list.

Learn more about pooled shRNA screening libraries.


shERWOOD-UltramiR shRNA Arrayed Whole Genome Library

shERWOOD-UltramiR shRNA arrayed library targets the human and mouse genomes. The library is provided as glycerol stock and contains approximately 19,000 genes with 4+ shRNA constructs per gene. 

Learn more about the development of library and shRNA designs.

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