Edit-R CRISPRa crRNA Libraries

Arrayed collections of synthetic CRISPR RNA for overexpression screening of gene families, druggable, and whole human genome.


Perform rapid gain-of-function studies with algorithm-designed CRISPR activation guide RNAs and dCas9-VPR expression

Edit-R CRISPRa libraries leverage the highly efficient two-component guide RNA system for gain-of-function arrayed studies using high-throughput, complex phenotypic analysis. Unlike lentiviral pooled screens, Edit-R synthetic crRNA arrayed libraries enable one-gene-per-well investigation using high-content assays to answer in-depth biological questions and enable studies for drug discovery, pathway analysis, or disease progression.

Don't see your gene family of choice? Upload your own gene to our Cherry-pick Library Plater for a fully customized CRISPRa screening library!

Highlights of Edit-R CRISPRa crRNA libraries

  • Available as four individual crRNAs per gene or a pool of four crRNAs to provide highly effective transcriptional activation (See Supporting Data tab)
  • crRNA pools provide robust activation while reducing costs, storage, and handling of libraries by 75% compared to working with four individual crRNAs
  • Algorithm-generated designs (published by Horlbeck, et. al.[1]) that demonstrate robust transcript activation (See References tab)
  • Chemically modified Edit-R crRNAs and tracrRNA provide additional stability against nuclease degradation and improve overall performance
  • Conveniently arrayed in 96- or 384-well plates and offered as gene family collections. Echo-qualified 384-well plates are available upon request

Required components for an Edit-R CRISPRa gene activation experiment using synthetic crRNA:

  • A lentiviral expression plasmid or lentiviral particles for dCas9-VPR[2]; a mammalian codon-optimized S. pyogenes deactivated Cas9 fused to VPR activation domains
    • Edit-R CRISPRa crRNA are also compatible with SunTag technology[3]
  • Edit-R trans-activating CRISPR RNA (tracrRNA)

If you're considering the purchase of a Druggable or Genome CRISPRa crRNA library, learn about how the Genomics Discovery Initiative can support your screening efforts!

Human Druggable is made up of: Proteases, Protein Kinases, Phosphatases, Transcription Factors, Ubiquitin Enzymes, GPCRs, Ion Channels and Drug Targets.

Human CRISPRa crRNA Libraries # genes (approximate) Catalog # (Pool/Set of 4)
Human Edit-R - Cell Cycle Regulation 169 GP/GC-103205-xx
Human Edit-R - Cytokine Receptors 110 GP/GC-104005-xx
Human Edit-R - Deubiquitinating Enzymes 94 GP/GC-104705-xx
Human Edit-R - DNA Damage Response 240 GP/GC-106005-xx
Human Edit-R - Drug Targets 3686 GP/GC-104650-xx
Human Edit-R - Druggable Genome 8422 GP/GC-104605-xx
Human Edit-R - Epigenetics 835 GP/GC-106105-xx
Human Edit-R - G Protein-coupled Receptors 390 GP/GC-103605-xx
Human Edit-R - Genome 19,127 GP/GC-105005-xx
Human Edit-R - Ion channels 417 GP/GC-103805-xx
Human Edit-R - Membrane Trafficking 140 GP/GC-105505-xx
Human Edit-R - Nuclear Receptors 52 GP/GC-103405-xx
Human Edit-R - Phosphatases 254 GP/GC-103705-xx
Human Edit-R - Proteases 576 GP/GC-105105-xx
Human Edit-R - Protein Kinases 746 GP/GC-103505-xx
Human Edit-R - Transcription Factors 1580 GP/GC-105805-xx
Human Edit-R - Tyrosine Kinases 90 GP/GC-103105-xx
Human Edit-R - Ubiquitin Enzymes 738 GP/GC-106205-xx
Mouse CRISPRa crRNA Libraries # genes (approximate) Catalog # (Pool/Set of 4)
Mouse Edit-R - Cell Cycle Regulation 105 GP/GC-113200-xx
Mouse Edit-R - Cytokine Receptors 139 GP/GC-114000-xx
Mouse Edit-R - Deubiquitinating Enzymes 100 GP/GC-114700-xx
Mouse Edit-R - Epigenetics 724 GP/GC-116100-xx
Mouse Edit-R - G Protein-coupled Receptors 515 GP/GC-113600-xx
Mouse Edit-R - Ion Channels 340 GP/GC-113800-xx
Mouse Edit-R - Membrane Trafficking 113 GP/GC-115505-xx
Mouse Edit-R - Nuclear Receptors 46 GP/GC-113400-xx
Mouse Edit-R - Phosphatases 273 GP/GC-113700-xx
Mouse Edit-R - Proteases 599 GP/GC-115100-xx
Mouse Edit-R - Protein Kinases 774 GP/GC-113500-xx
Mouse Edit-R - Transcription Factors 1419 GP/GC-115800-xx
Mouse Edit-R - Tyrosine Kinases 92 GP/GC-113105-xx
Mouse Edit-R - Ubiquitin Enzymes 752 GP/GC-116200-xx
  
HazardousNo
Shelf Life12 Months
Shipping ConditionAmbient
Stability at Recommended Storage ConditionsAt least 12 months
Storage Condition-20 C
crispra-crrna-individuals-pools-activation

Pooling of synthetic crRNAs can enhance transcriptional activation

crispra-crrna-individuals-pools-activation

Individual Edit-R CRISPRa crRNAs achieve robust target gene activation on their own, but when pooled together in a single reagent, enhanced activation levels can be achieved. U2OS cells stably expressing integrated dCas9-VPR were plated at 10,000 cells/well and transfected using DharmaFECT 4 Transfection Reagent with synthetic crRNA:tracrRNA targeting EGFR or POU5F1. The pre-designed crRNAs were used either individually or pooled (to a total concentration of 25 nM). Cells were harvested 72 hours post-transfection and the relative gene expression was measured using RT-qPCR. The relative fold transcriptional activation for each gene was calculated with the ∆∆Cq method using GAPDH as the reference gene and normalized to a non-targeting control (NTC).


dose-curve-synthetic-crrna-indiv-pools-crispra

Edit-R CRISPRa crRNAs and pools are highly effective at 25 nM working concentration

dose-curve-synthetic-crrna-indiv-pools-crispra

A dose curve of Edit-R CRISPRa crRNA:tracrRNA targeting two different genes demonstrates that a working concentration of 25 nM achieves robust target gene activation. U2OS cells stably expressing integrated dCas9-VPR were plated at 10,000 cells/well and transfected using DharmaFECT 4 Transfection Reagent with synthetic crRNA:tracrRNA targeting EGFR or POU5F1. The pre-designed crRNAs were used either individually or pooled at four concentrations (1, 5, 25, 100 nM). Cells were harvested 72 hours post-transfection and the relative gene expression was calculated using RT-qPCR. The relative expression of each gene was calculated with the ∆∆Cq method using GAPDH as the reference gene and normalized to a non-targeting control.


crispra-synthetic-crRNA-dCas9-VPR-stable-ttn-pou5f1

Efficient transcriptional gene activation with synthetic crRNA in multiple dCas9-VPR stable cells

crispra-synthetic-crRNA-dCas9-VPR-stable-ttn-pou5f1

Efficient transcriptional gene activation with synthetic crRNA:tracrRNA in dCas9-VPR stable cells. HEK293T, U2OS, MCF 10A, NIH/3T3 stably expressing integrated dCas9-VPR were plated at 10,000 cells/well and transfected using DharmaFECT Transfection Reagents with synthetic crRNA:tracrRNA (25 nM) targeting POU5F1 and TTN. K562 cells were electroporated with synthetic crRNA:tracrRNA (400 nM) targeting POU5F1 and TTN. Cells were harvested 72 hours post-transfection and the relative gene expression was measured using RT-qPCR. The relative fold transcriptional activation for each gene was calculated with the ∆∆Cq method using GAPDH as the reference gene and normalized to a non-targeting control.


crispra-timecourse-crrna-tracrrna

CRISPRa gene activation in U2OS cells is observed at 24 hours and peaks at 48-72 hours

crispra-timecourse-crrna-tracrrna

Edit-R CRISPRa synthetic crRNAs and pools achieve maximal activation at 48-72 hours post-transfection in dCas9-VPR-expressing cells. U2OS cells stably expressing integrated dCas9-VPR were plated at 10,000 cells/well and transfected using DharmaFECT 4 Transfection Reagent with synthetic crRNA:tracrRNA targeting EGFR, IL1R2, POU5F1 or TFAP2C. The four pre-designed crRNAs for CRISPRa were used either individually or pooled (to a total concentration of 25 nM). Cells were harvested at 24, 48, and 72 hours post-transfection and the relative gene expression was measured using qRT-PCR. The relative expression of each gene was calculated with the ∆∆Cq method using GAPDH as the reference gene and normalized to a non-targeting control.


References

  1. Horlbeck MA, Gilbert LA, et. al., Compact and highly active next-generation libraries for CRISPR-mediated gene repression and activation. 2016 Sep 23;5. pii: e19760. doi: 10.7554/eLife.19760. PubMed 27661255.

  2. Chavez A, Scheiman J et. al., Highly efficient Cas9-mediated transcriptional programming Nat Methods. 2015 Mar 2. doi: 10.1038/nmeth.3312. 10.1038/nmeth.3312 PubMed 2573049.

  3. Tanenbaum ME, Gilbert LA, et. al., A protein tagging system for signal amplification in gene expression and fluorescence imaging. Cell. 2014;159(3):635-646. doi:10.1016/j.cell.2014.09.039. PubMed 25307933