HB-EGF CRISPR Activation Plasmid (h): sc-400795-ACT

HBEGF encodes the heparin-binding EGF-like growth factor (HB-EGF), a membrane-anchored ligand that can be proteolytically shed to generate a soluble factor engaging EGFR and ERBB4. HB-EGF signaling activates MAPK/ERK, PI3K–AKT, and related transcriptional programs that regulate cell proliferation, survival, migration, and epithelial–mesenchymal interactions. It is implicated in wound repair, vascular remodeling, and inflammatory signaling through context-dependent crosstalk with ADAM metalloproteases and extracellular matrix components. Dysregulated HBEGF expression and EGFR-axis activation are frequently studied in cancer biology, fibrosis, and cardiovascular pathophysiology as mechanisms shaping cell-state transitions and microenvironmental responses.

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

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

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