Unbiased, powerful loss-of-function screens. The Edit-R CRISPR-Cas9 platform includes predesigned, ready-to-use guide RNAs. Achieve rapid assessment of multiple targets across many genes.
- Simplify CRISPR editing in all cell types including primary cells with Edit-R synthetic sgRNA libraries
- One-part guide format to carry out a do-it-yourself screening, avoids extra liquid handling which is created using the two-part system
- Guaranteed successful editing at the target site using proprietary Edit-R design algorithm
- Allows customers to perform larger screens
- More robust and reliable gene knockouts. Edit-R predesigned crRNA are selected by the Edit-R algorithm to be highly functional and specific to their target sequences
- Obtain high-confidence phenotypic results. Four unique crRNA designs per gene, provided as individuals or a pooled reagents
- Choose from arrayed 96-well or 384-well plates, available as gene family collections. Echo-qualified 384-well plates available upon request
- Flexibility and breadth. Also available as crRNA cherry-pick libraries; simply upload your own gene list and customize your plates
- Achieve high target sequence specificity and functionality for more robust, reliable gene knockouts using Edit-R lentiviral sgRNA, selected by the Edit-R algorithm
- Get multiple data points for increased experimental confidence and stratification of results by receiving four unique sgRNA designs per gene
- Choice and convenience with arrayed 96-well plates of E. coli glycerol stocks, also available as gene family collections
- To deal with more difficult cell types, plasmids can be isolated and delivered directly to cells or packaged into lentiviral particles
- Guarantee specificity with algorithm-designed lentiviral sgRNA pools targeting hundreds or thousands of genes supplied as high-titer, concentrated lentiviral particles
- Increase hit confidence and comprehensive genome screening by using 5 to 10 sgRNAs per gene
- Facilitate inter- and intra-pool normalization with over 400 positive and negative sgRNA controls in each pool
- Perform quality screens with increased hit confidence and greater biological reproducibility. Enough lentiviral particle volumes provided to have high average sgRNA fold representation
We're here to support you choose the right CRISPR-Cas9 screening platform to suit your scientific needs. Please refer to the following table for help:
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Lentiviral pooled sgRNA libraries
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Synthetic sgRNA/crRNA libraries
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Lentiviral arrayed sgRNA libraries
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Format
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Lentiviral particles in a small number of tubes
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Synthetic sgRNA/crRNA arrayed in multi-well plates
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Glycerol stocks containing lentiviral expression plasmids arrayed in 96-well plates
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Delivery to cells
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Lentiviral transduction
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Standard transfection reagents or electroporation
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Standard transfection reagents, electroporation, or lentiviral transduction if packaged into particles
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Assay types supported
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Phenotypes that result in changes in sgRNA abundance in the cell population and can be assessed by NGS can be investigated. Growth phenotypes (proliferation or survival) Selectable by cell sorting (fluorescence or cell surface marker expression)
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A wide range of cellular phenotypes (high content analysis, reporter or enzymatic assays, etc.) can be investigated due to one-gene-one-well layout. Editing efficiency in the population must be sufficient to allow for observation of the specific phenotype.
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A wide range of cellular phenotypes (high content analysis, reporter or enzymatic assays, etc.) can be investigated due to one-gene-one-well layout. Editing efficiency in the population must be sufficient to allow for observation of the specific phenotype. Phenotypes that result in changes in sgRNA abundance in the cell population and can be assessed by NGS and can be investigated if vectors are pooled
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Screening 'hands-on' time
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Relatively low; can be carried out in a single culture dish
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Scales up with the number of genes
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Scales up with the number of genes
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Data analysis requirements
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Next-gen sequencing required for identification of sgRNA(s) and its abundance in the cell population
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Phenotype is analyzed directly on a one-gene-per-well basis
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Phenotype is analyzed directly on a one-gene-per-well basis
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Ensure library specificity - algorithm-optimized guide RNA used in Edit-R crRNA and pooled sgRNA screening libraries.
Increase of mitotic index upon knockout of PLK1 and KIF11 with synthetic crRNA:tracrRNA
U2OS-(Ubi)EGFP-Cas9 stable cells were seeded at 5,000 cells/well in 96-well format. At 24 hours cells were transfected with 25 nM crRNA:tracrRNA complexes targeting the PLK1 and KIF11 and non-targeting controls (NTC) using 0.2 µl /well DharmaFECT4. Four different gene-targeting and NTC guide sequences were utilized and experimental controls included untransfected cells (UT) and cells treated with lipid alone. At 48 hours post-transfection, the phenotype was analyzed on the IN Cell Analyzer 2200 imaging system (GE Healthcare).
Upper panel: Mitotic Index (MI) was analyzed as percent cells positive for Phospho-Histone h6 (Ph6) and was normalized to the average MI of the four negative NTC crRNAs.
Lower panel: Representative fluorescent micrograph showing the nuclei in blue and Ph6 in red.
Algorithm applies to both synthetic crRNAs and expressed sgRNAs
Here we are targeting the gene PSMD11 at 12 different sites and measuring functionality by EGFP signal to indicate disruption of the proteasome. Cells were either transfected with crRNA:tracrRNAs, or transduced with lentiviral particles for expression of a sgRNA of the same design. There is no significant difference between the two guide RNA formats when targeting the same genomic site; a good design for crRNA will translate into a good design for sgRNA.
