Further your functional genomics studies with CRISPR-Cas9 gene editing using Edit-RTM crRNA libraries. Analyze hundreds of genes with multiple target sites per gene. Collections include human druggable and whole genome libraries.
Arrayed library format permits single-well analysis with many phenotypic assays; including high-content assays and other morphological or reporter assays.
Please note that Edit-R synthetic tracrRNATM is required for use with all synthetic crRNA products
Highlights of Edit-RTM crRNA libraries
- Selected by the Edit-RTM algorithm. Highly functional and target sequences specific for robust, reliable gene knockout. Chemically modified for improved nuclease resistance
- For high-confidence phenotypic results, provided as four unique crRNA designs per gene, as individuals or a pooled reagent.
- Conveniently arrayed in 96-or 384-well plates and offered as gene family collections. Echo-qualified 384-well plates are available upon request
- Also available as crRNA cherry-pick libraries; simply upload your own gene list and customize your plate
Human Druggable is made up of: Proteases, Protein Kinases, Phosphatases, Transcription Factors, Ubiquitin Enzymes, GPCRs, Ion Channels and Drug Targets.
|Human crRNA Libraries||# genes (approximate)||Catalog # (Pool/Set of 4)|
|Human Edit-RTM - Cell Cycle Regulation||169||GP/GC-003205-xx|
|Human Edit-RTM - Cytokine Receptors||110||GP/GC-004005-xx|
|Human Edit-RTM - Deubiquitinating Enzymes||94||GP/GC-004705-xx|
|Human Edit-RTM - DNA Damage Response||240||GP/GC-006005-xx|
|Human Edit-RTM - Drug Targets||3686||GP/GC-004650-xx|
|Human Edit-RTM - Druggable Genome||8422||GP/GC-004605-xx|
|Human Edit-RTM - Epigenetics||835||GP/GC-006105-xx|
|Human Edit-RTM - G Protein-coupled Receptors||390||GP/GC-003605-xx|
|Human Edit-RTM - Genome||19,127||GP/GC-005005-xx|
|Human Edit-RTM - Ion channels||417||GP/GC-003805-xx|
|Human Edit-RTM - Membrane Trafficking||140||GP/GC-005505-xx|
|Human Edit-RTM - Nuclear Receptors||52||GP/GC-003405-xx|
|Human Edit-RTM - Phosphatases||248||GP/GC-003705-xx|
|Human Edit-RTM - Proteases||527||GP/GC-005105-xx|
|Human Edit-RTM - Protein Kinases||746||GP/GC-003505-xx|
|Human Edit-RTM - Transcription Factors||1580||GP/GC-005805-xx|
|Human Edit-RTM - Tyrosine Kinases||90||GP/GC-003105-xx|
|Human Edit-RTM - Ubiquitin Enzymes||738||GP/GC-006205-xx|
|Mouse crRNA Libraries||# genes (approximate)||Catalog # (Pool/Set of 4)|
|Mouse Edit-RTM - Cell Cycle Regulation||105||GP/GC-013200-xx|
|Mouse Edit-RTM - Cytokine Receptors||139||GP/GC-014000-xx|
|Mouse Edit-RTM - Deubiquitinating Enzymes||100||GP/GC-014700-xx|
|Mouse Edit-RTM - Epigenetics||724||GP/GC-016100-xx|
|Mouse Edit-RTM - GProtein-coupled Receptors||515||GP/GC-013600-xx|
|Mouse Edit-RTM - Ion Channels||340||GP/GC-013800-xx|
|Mouse Edit-RTM - Membrane Trafficking||113||GP/GC-015505-xx|
|Mouse Edit-RTM - Nuclear Receptors||46||GP/GC-013400-xx|
|Mouse Edit-RTM - Phosphatases||273||GP/GC-013700-xx|
|Mouse Edit-RTM - Proteases||599||GP/GC-015100-xx|
|Mouse Edit-RTM - Protein Kinases||774||GP/GC-013500-xx|
|Mouse Edit-RTM - Transcription Factors||1419||GP/GC-015800-xx|
|Mouse Edit-R - Tyrosine Kinases||92||GP/GC-013105-xx|
|Mouse Edit-RTM - Ubiquitin Enzymes||752||GP/GC-016200-xx|
Increase of mitotic index upon knockout of PLK1 and KIF11 with synthetic crRNA:tracrRNA
U2OS-(Ubi)EGFP-Cas9 stable cells seeded at 5,000 cells/well in 96-well format were transfected the following day with four different crRNA:tracrRNA complexes at 25 nM concentration targeting PLK1 and KIF11. Four non-targeting crRNA controls (NTC), cells transfected with lipid alone (Lipid) or left untreated (UT) were used as negative controls. Cells were fixed at 48 hrs post-transfection and stained with anti Phospho-Histone h6 antibody (DY550 secondary antibody) and Hoechst 33342. Cell images were analyzed on the IN Cell Analyzer 2200 imaging system (GE Healthcare) and percent cells positive for Phospho-Histone h6 (Mitotic Index, MI) was normalized to the average MI of the negative NTC crRNAs.
Increase of cells stained with mitotic marker upon PLK1 and KIF11 knockout by synthetic crRNA:tracrRNA
U2OS-(Ubi)EGFP-Cas9 stable cells were plated at 5,000 cells/well in 96-well format and transfected the next day with 25 nM crRNA:tracrRNA targeting PLK1 or KIF11. Non-targeting crRNA (NTC1) or cell treated with lipid alone were used as negative controls. Cells were fixed at 48 hours post-transfection and stained with anti-Phosho-Histone h6 (Ph6) antibody (DY550 secondary antibody) and Hoechst 33342. Cell images were taken with IN Cell Analyzer 2200 imaging system (GE Healthcare). Fluorescent images presented with merged signal from nuclei (blue) and Ph6 (red).
Knockout of essential genes by synthetic crRNA:tracrRNA results in apoptosis
U2OS-(Ubi)EGFP-Cas9 stable cell seeded at 10,000 cells/well in 96-well format were transfected the following day with four different crRNA:tracrRNA complexes at 25 nM concentration targeting PLK1, KIF11 or BCL2L1 or four non-targeting crRNA controls (NTC). Wells transfected with lipid alone (Lipid) or left untreated (UT) were also included as controls. The effects on apoptosis were assayed using Casp3/9 homogeneous assay (ApoONE, Promega) at 48 hours post-transfection. Data normalized to average of NTC (non-targeting) crRNA controls.
Cell death phenotype observed following knockout of PLK1 and KIF11 with synthetic crRNA:tracrRNA
U2OS-(Ubi)EGFP-Cas9 stable cell seeded at 10,000 cells/well in 96-well format were transfected the following day with four different crRNA:tracrRNA complexes targeting PLK1 and KIF11 at 25 nM concentration. Non-targeting crRNA #1 (NTC-1) or PPIB crRNA were used as negative controls. Bright field images were taken at 48 hours post-transfection (20x, Leica DMIL microscope).
Gene knockout workflow using Cas9-expressing cells with synthetic crRNA:tracrRNA
For optimal results in a crRNA library workflow, it is recommended to establish stable expression of the Cas9 nuclease to improve knockout efficiency. Subsequent transfection of crRNA:tracrRNA is very straightforward and can be carried out in a high-throughput manner.
Ready-to-use, RNase-free Tris-HCl buffer for resuspension of synthetic crRNA and tracrRNA molecules