Erg-1/2/3 CRISPR Activation Plasmid (h): sc-400404-ACT

ERG encodes an ETS family transcription factor (Erg-1/2/3) that binds purine-rich DNA motifs to regulate gene programs controlling hematopoietic development, endothelial differentiation, and cell fate decisions. ERG integrates signals from MAPK/ERK and PI3K-associated pathways to coordinate transcriptional networks involved in proliferation, adhesion, and vascular homeostasis. Dysregulated ERG expression or rearrangement is frequently studied in oncogenic transcriptional rewiring, including leukemias and prostate cancer, and is also relevant to angiogenesis and inflammatory vascular biology. These features make ERG a widely used node for dissecting lineage-specifying transcriptional circuits and disease-associated gene regulatory networks.

Erg-1/2/3 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous ERG expression without altering the underlying DNA sequence.

Erg-1/2/3 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the ERG locus in human cell lines. The system is built around a catalytically inactive Cas9 (dCas9) carrying two inactivating mutations (D10A and N863A) that eliminate nuclease activity while preserving DNA binding. This dCas9 is fused to VP64, a potent transcriptional activator, and is co-expressed with a blasticidin resistance gene for selection. The second plasmid encodes the MS2-p65-HSF1 fusion protein, a secondary activator complex that works in concert with dCas9-VP64, alongside a hygromycin resistance gene. The third plasmid encodes a target-specific 20 nt sgRNA fused to two MS2 RNA aptamers that recruit the MS2-p65-HSF1 complex to the activation site, accompanied by a puromycin resistance gene. The three plasmids are delivered at a 1:1:1 mass ratio for balanced expression of all system components.

Once assembled at the target locus, the SAM complex binds within approximately 200 bp upstream of the ERG transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous Erg-1/2/3 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native ERG locus and enabling the study of Erg-1/2/3-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of Erg-1/2/3 pathway restoration in tumor cells with silenced or reduced ERG expression.

For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.