Erg CRISPR Activation Plasmid (m): sc-420226-ACT

Mouse Erg encodes an ETS family transcription factor that binds GGAA/T core motifs to regulate gene programs controlling endothelial specification, vascular stability, and hematopoietic stem and progenitor cell maintenance. Erg integrates signals from angiogenic and cytokine-responsive pathways, including VEGF-driven endothelial transcriptional networks and MAPK/PI3K-linked regulation of transcriptional output. Dysregulated ERG activity is widely studied in oncogenic transcriptional circuitry, particularly in hematologic malignancies and in ETS-driven fusion contexts, where altered target gene expression impacts proliferation, differentiation, and cell identity. In mouse systems, Erg is frequently used to model lineage commitment, vascular biology, and transcription factor dependency in normal development and disease-relevant cell states.

Erg CRISPR Activation Plasmid (m) provides a targeted, non-destructive approach to upregulating endogenous Erg expression without altering the underlying DNA sequence.

Erg CRISPR Activation Plasmid (m) 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 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-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of Erg pathway restoration in tumor cells with silenced or reduced Erg expression.

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