Edit-R CRISPRa crRNA Non-targeting Controls


Edit-R CRISPRa crRNA Non-targeting Controls are designed and recommended for use as negative controls for transcriptional activation experiments using Edit-R CRISPRa crRNA individuals and pools. These non-targeting controls will hybridize with tracrRNA and engage the dCas9-VPR complex, but will not target any PAM-adjacent sites in the human or mouse genome. Any observed alteration in gene expression levels or viability in cells treated with these controls can be used as a baseline response of the cells to the dCas9-VPR complex for comparison to those treated with target-specific CRISPRa crRNAs and pools.

Highlights

  • Proprietary alignment tools used to verify at least three mismatches or gaps to any potential target in human or mouse genomes
  • Choose an individual or pooled crRNA control to match your experimental crRNA reagent

Remember to order tracrRNA for use with your controls!

  
HazardousNo
Shipping ConditionAmbient
Stability at Recommended Storage ConditionsAt least 12 months
Storage Condition-20 C
Edit-R-CRISPRa-workflow-diagram

Edit-R CRISPRa workflow diagram with stable dCas9-VPR expression

Edit-R-CRISPRa-workflow-diagram

CRISPR activation workflow with lentiviral dCas9-VPR and synthetic crRNA:tracrRNA (left) or Lentiviral expressed sgRNA (right) as a purified lentiviral particle or plasmid prepared from glycerol stock.


crispra-crrna-individuals-pools-activation

Pooling of synthetic crRNAs can enhance transcriptional activation

crispra-crrna-individuals-pools-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 (25 nM) 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 calculated using RT-qPCR. The relative expression of each gene was calculated with the ∆∆Cq method using GAPDH as the housekeeping gene and normalized to a non-targeting control.