CRISPRa crRNA Libraries
Ensure library specificity - algorithm optimized guide RNA
Arrayed library collections of synthetic CRISPR crRNA for overexpression screening of gene families, druggable, and whole human genome. Perform rapid gain-of-function studies with CRISPR activation guide RNAs and dCas9-VPR expression.
Edit-R synthetic crRNA arrayed libraries enable one-gene-per-well investigation using high-content assays to answer in-depth biological questions. Further your drug discovery, pathway analysis, or disease progression studies.
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 compared to working with four individual crRNAs
- Algorithm-generated designs (published by Horlbeck, et. al.[1]) 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)
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 |
Shipping Condition | Ambient | |
Storage Conditions | -20 C | |
Stability at Recommended Storage Conditions | At least 12 months | |
Hazardous | No |
Pooling of synthetic crRNAs can enhance transcriptional 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).
Edit-R CRISPRa crRNAs and pools are highly effective at 25 nM working concentration
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.
Efficient transcriptional gene activation with synthetic crRNA in multiple dCas9-VPR stable cells
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 gene activation in U2OS cells is observed at 24 hours and peaks at 48-72 hours
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 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.
- 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.
-
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.
-
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