HE4 CRISPR Activation Plasmid (h): sc-402876-ACT

WFDC2 encodes HE4 (human epididymis protein 4), a secreted whey acidic protein (WAP) domain–containing glycoprotein implicated in extracellular protease regulation and remodeling of the pericellular microenvironment. HE4 expression is linked to epithelial differentiation programs and can influence cell–cell interactions, adhesion, and proteostasis within the secretory pathway and extracellular space. Dysregulated WFDC2/HE4 levels have been associated with altered tissue homeostasis and inflammatory signaling contexts in multiple epithelial-derived disease settings. As a circulating and tumor-associated protein in several malignancy-associated expression profiles, HE4 is frequently used as a molecular readout for pathway activation and phenotype stratification in translational research.

HE4 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous WFDC2 expression without altering the underlying DNA sequence.

HE4 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the WFDC2 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 WFDC2 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous HE4 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native WFDC2 locus and enabling the study of HE4-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of HE4 pathway restoration in tumor cells with silenced or reduced WFDC2 expression.

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