CRISPRmod CRISPRa synthetic crRNA libraries

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CRISPRmod CRISPRa synthetic crRNA libraries

CRISPRmod CRISPRa synthetic crRNA libraries

Algorithm optimized guide RNA design ensures specificity and functionality

Perform rapid gain-of-function screening with arrayed CRISPR activation guide RNA libraries. Find predefined collections for pathway, druggable, gene family or whole genome screening.

CRISPRmod CRISPRa 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 with CRISPR activation screening.

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 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 CRISPRa 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 CRISPRa gene activation experiment using synthetic crRNA:

A lentiviral expression plasmid or lentiviral particles for dCas9-VPR²; a mammalian codon-optimized S. pyogenes deactivated Cas9 fused to VPR activation domains
- CRISPRa crRNA are also compatible with SunTag technology³
trans-activating CRISPR RNA (tracrRNA)

Human CRISPRa crRNA Libraries	# genes (approximate)	Catalog # (Pool/Set of 4)
Cell Cycle Regulation	169	GP/GC-103205-xx
Cytokine Receptors	110	GP/GC-104005-xx
Deubiquitinating Enzymes	94	GP/GC-104705-xx
DNA Damage Response	240	GP/GC-106005-xx
Drug Targets	3686	GP/GC-104650-xx
Druggable Genome*	8422	GP/GC-104605-xx
Epigenetics	835	GP/GC-106105-xx
G Protein-coupled Receptors	390	GP/GC-103605-xx
Genome	19,127	GP/GC-105005-xx
Ion channels	417	GP/GC-103805-xx
Membrane Trafficking	140	GP/GC-105505-xx
Nuclear Receptors	52	GP/GC-103405-xx
Phosphatases	254	GP/GC-103705-xx
Proteases	576	GP/GC-105105-xx
Protein Kinases	746	GP/GC-103505-xx
Transcription Factors	1580	GP/GC-105805-xx
Tyrosine Kinases	90	GP/GC-103105-xx
Ubiquitin Enzymes	738	GP/GC-106205-xx

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

Efficient transcriptional gene activation with synthetic crRNA in multiple 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 CRISPRa synthetic crRNA:tracrRNA (25 nM) targeting POU5F1 and TTN. K562 cells were electroporated with CRISPRa 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

U2OS cells stably expressing integrated dCas9-VPR were plated at 10,000 cells/well and transfected using DharmaFECT 4 Transfection Reagent with CRISPRa synthetic crRNA:tracrRNA targeting EGFR, IL1R2, POU5F1 or TFAP2C. Four CRISPRa guides 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.

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

A CRISPRa dose curve 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 CRISPRa 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.