ERM CRISPR Activation Plasmid (h): sc-401161-ACT

Human ETV5 encodes the ETS family transcription factor ERM, a sequence-specific DNA-binding regulator that integrates extracellular cues to control gene programs governing proliferation, differentiation, and cell migration. ERM functions downstream of receptor tyrosine kinase signaling, including FGF–MAPK/ERK pathways, and contributes to lineage specification and tissue remodeling through transcriptional control of developmental and stem-like states. Dysregulated ETV5/ERM activity has been associated with altered epithelial and endocrine differentiation and is frequently linked to oncogenic transcriptional networks in multiple tumor contexts. These properties make ETV5 a useful node for studying ETS-driven transcriptional circuitry, signal-dependent enhancer usage, and context-specific gene regulation in human cell models.

ERM CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous ETV5 expression without altering the underlying DNA sequence.

ERM CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the ETV5 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 ETV5 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous ERM expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native ETV5 locus and enabling the study of ERM-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of ERM pathway restoration in tumor cells with silenced or reduced ETV5 expression.

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