ERF CRISPR Activation Plasmid (h): sc-403877-ACT

ERF (ETS2 repressor factor) is a human ETS-family transcriptional repressor that binds ETS motifs to modulate gene programs controlling cell-cycle progression, differentiation, and apoptosis. ERF activity is regulated by MAPK/ERK-dependent phosphorylation, which influences its subcellular localization and transcriptional output, linking extracellular growth signals to nuclear transcriptional control. By constraining ETS-driven transcription, ERF contributes to homeostatic regulation of proliferation and lineage specification, processes frequently perturbed in oncogenic signaling contexts. Dysregulation of ERF-dependent networks has been associated with altered growth factor responsiveness and aberrant transcriptional states relevant to cancer biology and developmental disorders.

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

ERF CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the ERF 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 ERF transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous ERF expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native ERF locus and enabling the study of ERF-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of ERF pathway restoration in tumor cells with silenced or reduced ERF expression.

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