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.

Predesigned CRISPRa guide RNA

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

 

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

Edit-R 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

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

  • CRISPRa positive crRNA controls

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

  • CRISPRa non-targeting crRNA controls

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

  • CRISPRa 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

Edit-R 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