CRISPRa for Transcriptional Activation

CRISPR activation (CRISPRa) reagents provide a straightforward, efficient set of tools to study a gene’s function by overexpression in its native context. The system requires two components to operate: a guide RNA and a catalytically deactivated Cas9 (dCas9)

Predesigned CRISPRa guide RNA

Algorithm-designed lentiviral and synthetic reagents specifically target promoter regions to induce a gene’s transcriptional activation.

CRISPRa dCas9-VPR

Nuclease-deactivated Cas9, fused to three transcriptional activation domains.

SAM transcriptional activation system

  • SAM tracrRNA

    For use with synthetic CRISPRa crRNA and a stable cell line expressing both components (NLS-dCas9-VP64 and MS2-p65-HSF1) of the SAM activation system.

CRISPR-Cas9 is not only for creating genomic double strand breaks, it can also be used to target promoter regions to activate or inhibit transcription. Using a deactivated or dead Cas9 nuclease fused to transcription activators, like dCas9-VPR, in combination with a guide RNA in either a synthetic or lentiviral format, the endogenous expression of a gene can be up-regulated by anywhere from 5 to 50,000+ fold!

CRISPRa assay considerations for reagent selection

dCas9-VPR source Guide RNA format Delivery method Recommendations for use & benefits
dCas9-VPR lentiviral particles Transduction & selection for dCas9-VPR stable cells Synthetic crRNA Transfection or electroporation
  • Short timepoint assays (2-4 days)
  • Ability to pool crRNAs for increased target gene activation
  • Multiplex to activate >1 gene
  • Arrayed screening
Lentiviral sgRNA particles Transduction
  • Extended timepoint assays with stable expression
  • Low MOI - single integration for stable and even activation in cell population
Lentiviral sgRNA plasmid Transfection or electroporation
  • Short timepoint assays
  • Puromycin selection to enrich
  • Multiplex to activate >1 gene
dCas9-VPR lentiviral plasmid Lentiviral sgRNA plasmid Co-transfection or electroporation with dCas9-VPR plasmid
  • Short timepoint assays
  • Antibiotics selection to enrich
  • No lentiviral integration
dCas9-VPR mRNA Synthetic crRNA Co-transfection or electroporation
  • Short timepoint assays
  • Antibiotic and fluorescent options to enrich
  • No exogenous DNA, no integration of reagents in genome

CRISPRa synthetic guide RNA

  • CRISPRa predesigned synthetic crRNA

    Offering coverage of the human and mouse genomes, these synthetic RNAs are modified for nuclease resistance and available as pooled or individual crRNA reagents.

  • Synthetic tracrRNA

    Trans-activating CRISPR RNA (tracrRNA) is a synthetic, HPLC-purified, long RNA required for use with crRNA to form the complex that programs dCas-VPR. It is modified for nuclease resistance.

  • CRISPRa synthetic crRNA positive controls

    Pooled or individual crRNA controls for assessment of optimal experimental conditions for gene activation

  • CRISPRa synthetic crRNA non-targeting controls

    Non-targeting controls to evaluate baseline cellular responses to CRISPRa components in the absence of gene target-specific crRNA

  • CRISPRa synthetic crRNA libraries

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

  • Cherry-pick libraries

    Upload your own gene list to customize and order plates of predesigned crRNA for CRISPRa studies across tens or thousands of genes

CRISPRa lentiviral guide RNA

CRISPRa products may be combined in several ways to support your particular experimental needs. Whether you’re doing co-transfection or creating a stable dCas9-VPR cell line (recommended), choose from our synthetic RNA, lentiviral particles, or glycerol stock (plasmid) sgRNA, and combine with dCas9-VPR plasmid or lentiviral particles.

Workflow for CRISPRa transcriptional activation with stably expressing dCas9-VPR cells
Workflow for CRISPRa transcriptional activation with stably expressing dCas9-VPR cells
CRISPRa co-transfection workflow
Edit-R CRISPRa co-transfection workflow