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CRISPRa/i sgRNA clones

Both CRISPRa and CRISPRi systems use a dCas9 construct fused to either activators or repressors to leverage the power of PAM-anchored targeting for transcriptional modulation. These systems simply bring the precision of CRISPR to gene modulation.
CRISPRa sgRNA

CRISPR activation sgRNA clones

Up-regulate endogenous gene expression, without cutting the DNA

Vector Promoter CRISPR option Selection Marker / Reporter Gene Vector Type Vector Map
pCRISPR-AG01 U6 CRISPRa dCas9 SAM Puro / eGFP Lentiviral get_info
pCRISPR-AG02 U6 CRISPRa dCas9 SAM Puro / eGFP Lentiviral get_info

Application

The CRISPR (SAM) system (CRISPR-based Synergistic Activation Mediator) is an astute combination of dCas9 and other molecular biology tools designed to activate the transcription of any endogenous gene of interest. The system comprises 3 components that form a DNA-binding complex once transfected into the cells.
  • dCas9 fused to transcriptional activator VP64
  • sgRNA with an MS2 adaptor forming a characteristic stem-loop structure
  • MCP-p65-HSF1 fusion protein (MPH complex)
 
dCas9-VP64, the MPH complex, and sgRNA were co-overexpressed in HEK293 cells. The transcriptional activity of VEGFA (Figure A), EGFR (Figure B), and HBG (Figure C) was detected by qPCR. The cell proliferation curves for cells transduced with sgRNA are shown in green, while those for cells without sgRNA transduction are shown in blue. The transcription levels of the target genes were compared with those of GAPDH, indicating a significant increase in the transcriptional levels of the target genes in HEK293 cells.

 

CRISPRa related products

CRISPRa controls
Controls Description Selection Marker / Reporter Gene Vector Type Vector Map
CCPCTR01-AG01 Scrambled sgRNA control for pCRISPR-AG01 Puro / eGFP Lentiviral get_info
CCPCTR01-AG02 Scrambled sgRNA control for pCRISPR-AG02 Puro / eGFP Lentiviral get_info
GeneHero™ CRISPRa Stable Cell Lines
  • The CRISPRa cell lines have stably integrated human codon-optimized dCas9 fused at the C-terminus of transcriptional activator VP64, and activation helper protein complex MPH (MS2-p65-HSF1).
CRISPR-Fectin™ Transfection Reagent
  • An optimized lipid-based transfection reagent for CRISPR-Cas9 ribonucleoprotein delivery